TY - JOUR AU - Szabó, Eszter AU - Nemes-Nikodém, Éva AU - Vass, Krisztina Rubina AU - Zámbó, Zsófia Melinda AU - Zrupko, E. AU - Törőcsik, Beáta AU - Ozohanics, Olivér AU - Nagy, Bálint AU - Ambrus, Attila TI - Structural and Biochemical Investigation of Selected Pathogenic Mutants of the Human Dihydrolipoamide Dehydrogenase JF - INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES J2 - INT J MOL SCI VL - 24 PY - 2023 IS - 13 PG - 23 SN - 1661-6596 DO - 10.3390/ijms241310826 UR - https://m2.mtmt.hu/api/publication/34075046 ID - 34075046 N1 - Export Date: 1 September 2023 Correspondence Address: Ambrus, A.; Department of Biochemistry, 37-47 Tuzolto St, Hungary; email: ambrus.attila@med.semmelweis-univ.hu Funding details: TKP2021-EGA-25, ÚNKP-19-3-III-SE-17 Funding details: Horizon 2020 Framework Programme, H2020, PID18322 Funding details: Helmholtz-Zentrum Berlin für Materialien und Energie, HZB Funding details: Semmelweis Egyetem, STIA-OTKA-2021 Funding details: Hungarian Scientific Research Fund, OTKA, 143627 Funding details: Helmholtz-Zentrum für Umweltforschung, UFZ Funding details: Nemzeti Kutatási, Fejlesztési és Innovaciós Alap, NKFIA Funding text 1: This research was funded by the Hungarian Brain Research Program 2 (2017-1.2.1-NKP-2017-00002 grant, to Vera Adam-Vizi), Semmelweis University (STIA-OTKA-2021 grant, to A.A.), Hungarian Scientific Research Fund (OTKA grant 143627, to A.A.), Ministry of Innovation and Technology of Hungary (TKP2021-EGA-25 grant, to A.A. and ÚNKP-19-3-III-SE-17 grant of the New National Excellence Program, to E.S.; project no. TKP2021-EGA-25 has been implemented with the support provided by the Ministry of Innovation and Technology of Hungary from the National Research, Development and Innovation Fund, financed under the TKP2021-EGA funding scheme), and European Union’s Horizon 2020 Research and Innovation Programme (Structural Biology Research Infrastructures for Translational Research and Discovery, iNEXT-Discovery PID18322 and MX-212-00269-ST (Helmholtz Zentrum Berlin/CALIPSO) grants, both to A.A.). Funding text 2: The authors gratefully acknowledge the generous support from Vera Adam-Vizi (Semmelweis University) and Manfred S. Weiss (Helmholtz-Zentrum Berlin). AB - Clinically relevant disease-causing variants of the human dihydrolipoamide dehydrogenase (hLADH, hE3), a common component of the mitochondrial α-keto acid dehydrogenase complexes, were characterized using a multipronged approach to unravel the molecular pathomechanisms that underlie hLADH deficiency. The G101del and M326V substitutions both reduced the protein stability and triggered the disassembly of the functional/obligate hLADH homodimer and significant FAD losses, which altogether eventually manifested in a virtually undetectable catalytic activity in both cases. The I12T-hLADH variant proved also to be quite unstable, but managed to retain the dimeric enzyme form; the LADH activity, both in the forward and reverse catalytic directions and the affinity for the prosthetic group FAD were both significantly compromised. None of the above three variants lent themselves to an in-depth structural analysis via X-ray crystallography due to inherent protein instability. Crystal structures at 2.89 and 2.44 Å resolutions were determined for the I318T- and I358T-hLADH variants, respectively; structure analysis revealed minor conformational perturbations, which correlated well with the residual LADH activities, in both cases. For the dimer interface variants G426E-, I445M-, and R447G-hLADH, enzyme activities and FAD loss were determined and compared against the previously published structural data. © 2023 by the authors. LA - English DB - MTMT ER - TY - JOUR AU - Tőkés, Anna-Mária AU - Vári-Kakas, S. AU - Kulka, Janina AU - Törőcsik, Beáta TI - Tumor Glucose and Fatty Acid Metabolism in the Context of Anthracycline and Taxane-Based (Neo)Adjuvant Chemotherapy in Breast Carcinomas JF - FRONTIERS IN ONCOLOGY J2 - FRONT ONCOL VL - 12 PY - 2022 PG - 23 SN - 2234-943X DO - 10.3389/fonc.2022.850401 UR - https://m2.mtmt.hu/api/publication/32804574 ID - 32804574 LA - English DB - MTMT ER - TY - JOUR AU - Mihályi, Csaba AU - Iordanov, Iordan AU - Törőcsik, Beáta AU - Csanády, László TI - Simple binding of protein kinase A, prior to phosphorylation, allows CFTR anion channels to be opened by nucleotides JF - PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA J2 - P NATL ACAD SCI USA VL - 117 PY - 2020 IS - 35 SP - 21740 EP - 21746 PG - 7 SN - 0027-8424 DO - 10.1073/pnas.2007910117 UR - https://m2.mtmt.hu/api/publication/31397038 ID - 31397038 LA - English DB - MTMT ER - TY - JOUR AU - Csanády, László AU - Törőcsik, Beáta TI - Cystic fibrosis drug ivacaftor stimulates CFTR channels at picomolar concentrations JF - ELIFE J2 - ELIFE VL - 8 PY - 2019 PG - 18 SN - 2050-084X DO - 10.7554/eLife.46450 UR - https://m2.mtmt.hu/api/publication/30775333 ID - 30775333 AB - The devastating inherited disease cystic fibrosis (CF) is caused by mutations of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) anion channel. The recent approval of the CFTR potentiator drug ivacaftor (Vx-770) for the treatment of CF patients has marked the advent of causative CF therapy. Currently, thousands of patients are being treated with the drug, and its molecular mechanism of action is under intensive investigation. Here we determine the solubility profile and true stimulatory potency of Vx-770 towards wild-type (WT) and mutant human CFTR channels in cell-free patches of membrane. We find that its aqueous solubility is ~200 fold lower (~60 nanomolar), whereas the potency of its stimulatory effect is >100 fold higher, than reported, and is unexpectedly fully reversible. Strong, but greatly delayed, channel activation by picomolar Vx-770 identifies multiple sequential slow steps in the activation pathway. These findings provide solid guidelines for the design of in vitro studies using Vx-770. © 2019, Csanády and Töröcsik. LA - English DB - MTMT ER - TY - JOUR AU - Szabó, Eszter AU - Wilk, Piotr AU - Nagy, Bálint AU - Zámbó, Zsófia Melinda AU - Bui, Dávid AU - Weichsel, Andrzej AU - Arjunan, Palaniappa AU - Törőcsik, Beáta AU - Hubert, Agnes AU - Furey, William AU - Montfort, William R AU - Jordan, Frank AU - Weiss, Manfred S AU - Ádám, Veronika AU - Ambrus, Attila TI - Underlying molecular alterations in human dihydrolipoamide dehydrogenase deficiency revealed by structural analyses of disease-causing enzyme variants JF - HUMAN MOLECULAR GENETICS J2 - HUM MOL GENET VL - 28 PY - 2019 IS - 20 SP - 3339 EP - 3354 PG - 16 SN - 0964-6906 DO - 10.1093/hmg/ddz177 UR - https://m2.mtmt.hu/api/publication/30799456 ID - 30799456 AB - Human dihydrolipoamide dehydrogenase (hLADH, hE3) deficiency (OMIM# 246900) is an often prematurely lethal genetic disease usually caused by inactive or partially inactive hE3 variants. Here we report the crystal structure of wild-type hE3 at an unprecedented high resolution of 1.75 Å and the structures of six disease-causing hE3 variants at resolutions ranging from 1.44 to 2.34 Å. P453L proved to be the most deleterious substitution in structure as aberrations extensively compromised the active site. The most prevalent G194C-hE3 variant primarily exhibited structural alterations close to the substitution site, whereas the nearby cofactor-binding residues were left unperturbed. The G426E substitution mainly interfered with the local charge distribution introducing dynamics to the substitution site in the dimer interface; G194C and G426E both led to minor structural changes. The R460G, R447G, and I445M substitutions all perturbed a solvent accessible channel, the so-called H+/H2O channel, leading to the active site. Molecular pathomechanisms of enhanced reactive oxygen species (ROS) generation and impaired binding to multienzyme complexes were also addressed according to the structural data for the relevant mutations. In summary, we present here for the first time a comprehensive study that links three-dimensional structures of disease-causing hE3 variants to residual hLADH activities, altered capacities for ROS generation, compromised affinities for multienzyme complexes, and eventually clinical symptoms. Our results may serve as useful starting points for future therapeutic intervention approaches. LA - English DB - MTMT ER - TY - JOUR AU - Zahola, Péter AU - Hanics, János AU - Pintér, Panka AU - Máté, Zoltán AU - Gáspárdy, Anna AU - Hevesi, Zsófia AU - Echevarria, Diego AU - Adori, Csaba AU - Barde, Swapnali AU - Törőcsik, Beáta AU - Erdélyi, Ferenc AU - Szabó, Gábor AU - Wagner, Ludwig AU - Kovács, Gábor Géza AU - Hökfelt, Tomas AU - Harkany, Tibor AU - Alpár, Alán TI - Secretagogin expression in the vertebrate brainstem with focus on the noradrenergic system and implications for Alzheimer's disease JF - BRAIN STRUCTURE & FUNCTION J2 - BRAIN STRUCT FUNC VL - 224 PY - 2019 IS - 6 SP - 2061 EP - 2078 PG - 18 SN - 1863-2653 DO - 10.1007/s00429-019-01886-w UR - https://m2.mtmt.hu/api/publication/30723780 ID - 30723780 N1 - SE NAP B Research Group of Experimental Neuroanatomy and Developmental Biology, Semmelweis University, Budapest, Hungary Department of Anatomy, Semmelweis University, Budapest, Hungary Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest, Hungary Department of Molecular Neurosciences, Center for Brain Research, Medical University of Vienna, Vienna, 1090, Austria Institute of Neuroscience, University of Miguel Hernandez de Elche, Alicante, Spain Department of Neuroscience, Karolinska Institutet, Biomedicum 7D, Stockholm, SE-17165, Sweden Department of Medical Biochemistry, Semmelweis University, Budapest, Hungary Department of Internal Medicine III, Medical University of Vienna, Vienna, Austria Institute of Neurology, Medical University of Vienna, Vienna, Austria Cited By :12 Export Date: 31 October 2023 Correspondence Address: Alpár, A.; SE NAP B Research Group of Experimental Neuroanatomy and Developmental Biology, Hungary; email: Alpar.Alan@med.semmelweis-univ.hu AB - Calcium-binding proteins are widely used to distinguish neuronal subsets in the brain. This study focuses on secretagogin, an EF-hand calcium sensor, to identify distinct neuronal populations in the brainstem of several vertebrate species. By using neural tube whole mounts of mouse embryos, we show that secretagogin is already expressed during the early ontogeny of brainstem noradrenaline cells. In adults, secretagogin-expressing neurons typically populate relay centres of special senses and vegetative regulatory centres of the medulla oblongata, pons and midbrain. Notably, secretagogin expression overlapped with the brainstem column of noradrenergic cell bodies, including the locus coeruleus (A6) and the A1, A5 and A7 fields. Secretagogin expression in avian, mouse, rat and human samples showed quasi-equivalent patterns, suggesting conservation throughout vertebrate phylogeny. We found reduced secretagogin expression in locus coeruleus from subjects with Alzheimer's disease, and this reduction paralleled the loss of tyrosine hydroxylase, the enzyme rate limiting noradrenaline synthesis. Residual secretagogin immunoreactivity was confined to small submembrane domains associated with initial aberrant tau phosphorylation. In conclusion, we provide evidence that secretagogin is a useful marker to distinguish neuronal subsets in the brainstem, conserved throughout several species, and its altered expression may reflect cellular dysfunction of locus coeruleus neurons in Alzheimer's disease. LA - English DB - MTMT ER - TY - JOUR AU - Szabó, Eszter AU - Mizsei, Réka AU - Wilk, P AU - Zámbó, Zsófia Melinda AU - Törőcsik, Beáta AU - Weiss, MS AU - Ádám, Veronika AU - Ambrus, Attila TI - Crystal structures of the disease-causing D444V mutant and the relevant wild type human dihydrolipoamide dehydrogenase JF - FREE RADICAL BIOLOGY AND MEDICINE J2 - FREE RADICAL BIO MED VL - 124 PY - 2018 SP - 214 EP - 220 PG - 7 SN - 0891-5849 DO - 10.1016/j.freeradbiomed.2018.06.008 UR - https://m2.mtmt.hu/api/publication/3407990 ID - 3407990 N1 - Department of Medical Biochemistry, MTA-SE Laboratory for Neurobiochemistry, Semmelweis University, Budapest, H-1094, Hungary Macromolecular Crystallography, Helmholtz-Zentrum Berlin für Materialien und Energie, D-12489, Berlin, Germany Cited By :2 Export Date: 24 January 2020 CODEN: FRBME Correspondence Address: Ambrus, A.; Department of Medical Biochemistry, Semmelweis University, 37-47 Tuzolto Street, Hungary; email: ambrus.attila@med.semmelweis-univ.hu Chemicals/CAS: dihydrolipoamide dehydrogenase, 37340-89-9, 9001-18-7; Dihydrolipoamide Dehydrogenase; Mutant Proteins Funding details: Hungarian Scientific Research Fund Funding details: Hungarian Scientific Research Fund, 112230 Funding details: Magyar Tudományos Akadémia, 02001 Funding text 1: Financial support was secured from the Hungarian Academy of Sciences (MTA grant 02001 to V.A-V.), the Hungarian Scientific Research Fund (OTKA grant 112230 to V.A-V.), the Hungarian Brain Research Program (grants KTIA_13_NAP-A-III/6. and 2017-1.2.1-NKP-2017-00002 to V.A-V.), the EMBO , Fulbright , and Bolyai Fellowships (to A.A.), and the Young Investigator Research Grants of the Semmelweis University and Gedeon Richter Pharmaceuticals PIc . (to A.A.). Data collection at Helmholtz-Zentrum Berlin was carried out under contract number 16204087-ST. Department of Medical Biochemistry, MTA-SE Laboratory for Neurobiochemistry, Semmelweis University, Budapest, H-1094, Hungary Macromolecular Crystallography, Helmholtz-Zentrum Berlin für Materialien und Energie, D-12489, Berlin, Germany Cited By :3 Export Date: 4 January 2021 CODEN: FRBME Correspondence Address: Ambrus, A.; Department of Medical Biochemistry, Semmelweis University, 37-47 Tuzolto Street, Hungary; email: ambrus.attila@med.semmelweis-univ.hu Chemicals/CAS: dihydrolipoamide dehydrogenase, 37340-89-9, 9001-18-7; Dihydrolipoamide Dehydrogenase; Mutant Proteins Funding details: Hungarian Scientific Research Fund, OTKA Funding details: Magyar Tudományos Akadémia, MTA Funding details: Hungarian Scientific Research Fund, OTKA, 112230 Funding details: Magyar Tudományos Akadémia, MTA, 02001 Funding text 1: Financial support was secured from the Hungarian Academy of Sciences (MTA grant 02001 to V.A-V.), the Hungarian Scientific Research Fund (OTKA grant 112230 to V.A-V.), the Hungarian Brain Research Program (grants KTIA_13_NAP-A-III/6. and 2017-1.2.1-NKP-2017-00002 to V.A-V.), the EMBO , Fulbright , and Bolyai Fellowships (to A.A.), and the Young Investigator Research Grants of the Semmelweis University and Gedeon Richter Pharmaceuticals PIc . (to A.A.). Data collection at Helmholtz-Zentrum Berlin was carried out under contract number 16204087-ST. Department of Medical Biochemistry, MTA-SE Laboratory for Neurobiochemistry, Semmelweis University, Budapest, H-1094, Hungary Macromolecular Crystallography, Helmholtz-Zentrum Berlin für Materialien und Energie, D-12489, Berlin, Germany Cited By :3 Export Date: 5 January 2021 CODEN: FRBME Correspondence Address: Ambrus, A.; Department of Medical Biochemistry, Semmelweis University, 37-47 Tuzolto Street, Hungary; email: ambrus.attila@med.semmelweis-univ.hu Chemicals/CAS: dihydrolipoamide dehydrogenase, 37340-89-9, 9001-18-7; Dihydrolipoamide Dehydrogenase; Mutant Proteins Funding details: Hungarian Scientific Research Fund, OTKA Funding details: Magyar Tudományos Akadémia, MTA Funding details: Hungarian Scientific Research Fund, OTKA, 112230 Funding details: Magyar Tudományos Akadémia, MTA, 02001 Funding text 1: Financial support was secured from the Hungarian Academy of Sciences (MTA grant 02001 to V.A-V.), the Hungarian Scientific Research Fund (OTKA grant 112230 to V.A-V.), the Hungarian Brain Research Program (grants KTIA_13_NAP-A-III/6. and 2017-1.2.1-NKP-2017-00002 to V.A-V.), the EMBO , Fulbright , and Bolyai Fellowships (to A.A.), and the Young Investigator Research Grants of the Semmelweis University and Gedeon Richter Pharmaceuticals PIc . (to A.A.). Data collection at Helmholtz-Zentrum Berlin was carried out under contract number 16204087-ST. AB - We report the crystal structures of the human (dihydro)lipoamide dehydrogenase (hLADH, hE3) and its disease-causing homodimer interface mutant D444V-hE3 at 2.27 and 1.84 Å resolution, respectively. The wild type structure is a unique uncomplexed, unliganded hE3 structure with the true canonical sequence. Based on the structural information a novel molecular pathomechanism is proposed for the impaired catalytic activity and enhanced capacity for reactive oxygen species generation of the pathogenic mutant. The mechanistic model involves a previously much ignored solvent accessible channel leading to the active site that might be perturbed also by other disease-causing homodimer interface substitutions of this enzyme. © 2018 Elsevier Inc. LA - English DB - MTMT ER - TY - JOUR AU - Sorum, Ben AU - Törőcsik, Beáta AU - Csanády, László TI - Asymmetry of movements in CFTR's two ATP sites during pore opening serves their distinct functions JF - ELIFE J2 - ELIFE VL - 6 PY - 2017 PG - 17 SN - 2050-084X DO - 10.7554/eLife.29013 UR - https://m2.mtmt.hu/api/publication/3287727 ID - 3287727 AB - CFTR, the chloride channel mutated in cystic fibrosis (CF) patients, is opened by ATP binding to two cytosolic nucleotide binding domains (NBDs), but pore-domain mutations may also impair gating. ATP-bound NBDs dimerize occluding two nucleotides at interfacial binding sites; one site hydrolyzes ATP, the other is inactive. The pore opens upon tightening, and closes upon disengagement, of the catalytic site following ATP hydrolysis. Extent, timing, and role of non-catalytic-site movements are unknown. Here we exploit equilibrium gating of a hydrolysis-deficient mutant and apply Phi value analysis to compare timing of opening-associated movements at multiple locations, from the cytoplasmic ATP sites to the extracellular surface. Marked asynchrony of motion in the two ATP sites reveals their distinct roles in channel gating. The results clarify the molecular mechanisms of functional cross-talk between canonical and degenerate ATP sites in asymmetric ABC proteins, and of the gating defects caused by two common CF mutations. LA - English DB - MTMT ER - TY - JOUR AU - Ambrus, Attila AU - Wang, J AU - Mizsei, Réka AU - Zámbó, Zsófia Melinda AU - Törőcsik, Beáta AU - Jordan, F AU - Ádám, Veronika TI - Structural alterations induced by ten disease-causing mutations of human dihydrolipoamide dehydrogenase analyzed by hydrogen/deuterium-exchange mass spectrometry: Implications for the structural basis of E3 deficiency JF - BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR BASIS OF DISEASE J2 - BBA-MOL BASIS DIS VL - 1862 PY - 2016 IS - 11 SP - 2098 EP - 2109 PG - 12 SN - 0925-4439 DO - 10.1016/j.bbadis.2016.08.013 UR - https://m2.mtmt.hu/api/publication/3114546 ID - 3114546 N1 - Attila Ambrus, Junjie Wang and Reka Mizsei contributed equally to this work. Department of Medical Biochemistry, MTA-SE Laboratory for Neurobiochemistry, Semmelweis University, Budapest, Hungary Department of Chemistry, Rutgers University, Newark, NJ, United States Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University, New York, NY, United States Laboratory of Immunobiology, Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, United States Cited By :9 Export Date: 28 May 2020 CODEN: BBADE Correspondence Address: Ambrus, A.; Department of Medical Biochemistry, MTA-SE Laboratory for Neurobiochemistry, Semmelweis UniversityHungary; email: ambrus.attila@med.semmelweis-univ.hu Chemicals/CAS: amino acid, 65072-01-7; deuterium, 7782-39-0; dihydrolipoamide dehydrogenase, 37340-89-9, 9001-18-7; hydrogen, 12385-13-6, 1333-74-0 Funding details: Magyar Tudományos Akadémia, MTA Funding details: Hungarian Scientific Research Fund, OTKA Funding details: Hungarian Scientific Research Fund, OTKA, 112230 Funding details: Magyar Tudományos Akadémia, MTA, 02001 Funding text 1: We are grateful to Drs. Oliver Ozohanics, Karoly Vekey (both from the Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest, Hungary) and Arpad Somogyi (Ohio State University, Columbus, OH, USA) for their contributions to the method development in mass spectrometry. Financial support is gratefully acknowledged from the Hungarian Academy of Sciences (MTA grant 02001 to V.A.-V.), the Hungarian Scientific Research Fund (OTKA, grant 112230 to V.A.-V.), the Hungarian Brain Research Program (grant KTIA_13_NAP-A-III/6 to V.A.-V.), the Bolyai and the Fulbright Fellowships (to A.A.), NIH - R15GM116077 and NSF-CHE - 1402675 (to F.J.). Department of Medical Biochemistry, MTA-SE Laboratory for Neurobiochemistry, Semmelweis University, Budapest, Hungary Department of Chemistry, Rutgers University, Newark, NJ, United States Laboratory of Mass Spectrometry and Gaseous Ion Chemistry, The Rockefeller University, New York, NY, United States Laboratory of Immunobiology, Department of Medical Oncology, Dana-Farber Cancer Institute, Boston, MA, United States Cited By :12 Export Date: 5 January 2021 CODEN: BBADE Correspondence Address: Ambrus, A.; Department of Medical Biochemistry, MTA-SE Laboratory for Neurobiochemistry, Semmelweis UniversityHungary; email: ambrus.attila@med.semmelweis-univ.hu Chemicals/CAS: amino acid, 65072-01-7; deuterium, 7782-39-0; dihydrolipoamide dehydrogenase, 37340-89-9, 9001-18-7; hydrogen, 12385-13-6, 1333-74-0 Funding details: Hungarian Scientific Research Fund, OTKA Funding details: Magyar Tudományos Akadémia, MTA Funding details: Hungarian Scientific Research Fund, OTKA, 112230 Funding details: Magyar Tudományos Akadémia, MTA, 02001 Funding text 1: We are grateful to Drs. Oliver Ozohanics, Karoly Vekey (both from the Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest, Hungary) and Arpad Somogyi (Ohio State University, Columbus, OH, USA) for their contributions to the method development in mass spectrometry. Financial support is gratefully acknowledged from the Hungarian Academy of Sciences (MTA grant 02001 to V.A.-V.), the Hungarian Scientific Research Fund (OTKA, grant 112230 to V.A.-V.), the Hungarian Brain Research Program (grant KTIA_13_NAP-A-III/6 to V.A.-V.), the Bolyai and the Fulbright Fellowships (to A.A.), NIH - R15GM116077 and NSF-CHE - 1402675 (to F.J.). AB - Pathogenic amino acid substitutions of the common E3 component (hE3) of the human alpha-ketoglutarate dehydrogenase and the pyruvate dehydrogenase complexes lead to severe metabolic diseases (E3 deficiency), which usually manifest themselves in cardiological and/or neurological symptoms and often cause premature death. To date, 14 disease-causing amino acid substitutions of the hE3 component have been reported in the clinical literature. None of the pathogenic protein variants has lent itself to high-resolution structure elucidation by X-ray or NMR. Hence, the structural alterations of the hE3 protein caused by the disease-causing mutations and leading to dysfunction, including the enhanced generation of reactive oxygen species by selected disease-causing variants, could only be speculated. Here we report results of an examination of the effects on the protein structure of ten pathogenic mutations of hE3 using hydrogen/deuterium-exchange mass spectrometry (HDX-MS), a new and state-of-the-art approach of solution structure elucidation. On the basis of the results, putative structural and mechanistic conclusions were drawn regarding the molecular pathogenesis of each disease-causing hE3 mutation addressed in this study. © 2016 Elsevier B.V. LA - English DB - MTMT ER - TY - JOUR AU - Dóczi, Judit AU - Törőcsik, Beáta AU - Echaniz-Laguna, A AU - Mousson, de Camaret B AU - Starkov, A AU - Starkova, N AU - Gál, Anikó AU - Molnár, Mária Judit AU - Kawamata, H AU - Manfredi, G AU - Ádám, Veronika AU - Chinopoulos, Christos TI - Alterations in voltage-sensing of the mitochondrial permeability transition pore in ANT1-deficient cells JF - SCIENTIFIC REPORTS J2 - SCI REP VL - 6 PY - 2016 PG - 21 SN - 2045-2322 DO - 10.1038/srep26700 UR - https://m2.mtmt.hu/api/publication/3069984 ID - 3069984 AB - The probability of mitochondrial permeability transition (mPT) pore opening is inversely related to the magnitude of the proton electrochemical gradient. The module conferring sensitivity of the pore to this gradient has not been identified. We investigated mPT's voltage-sensing properties elicited by calcimycin or H2O2 in human fibroblasts exhibiting partial or complete lack of ANT1 and in C2C12 myotubes with knocked-down ANT1 expression. mPT onset was assessed by measuring in situ mitochondrial volume using the 'thinness ratio' and the 'cobalt-calcein' technique. De-energization hastened calcimycin-induced swelling in control and partially-expressing ANT1 fibroblasts, but not in cells lacking ANT1, despite greater losses of mitochondrial membrane potential. Matrix Ca(2+) levels measured by X-rhod-1 or mitochondrially-targeted ratiometric biosensor 4mtD3cpv, or ADP-ATP exchange rates did not differ among cell types. ANT1-null fibroblasts were also resistant to H2O2-induced mitochondrial swelling. Permeabilized C2C12 myotubes with knocked-down ANT1 exhibited higher calcium uptake capacity and voltage-thresholds of mPT opening inferred from cytochrome c release, but intact cells showed no differences in calcimycin-induced onset of mPT, irrespective of energization and ANT1 expression, albeit the number of cells undergoing mPT increased less significantly upon chemically-induced hypoxia than control cells. We conclude that ANT1 confers sensitivity of the pore to the electrochemical gradient. LA - English DB - MTMT ER - TY - JOUR AU - Mihályi, Csaba AU - Törőcsik, Beáta AU - Csanády, László TI - Obligate coupling of CFTR pore opening to tight nucleotide-binding domain dimerization JF - ELIFE J2 - ELIFE VL - 5 PY - 2016 PG - 12 SN - 2050-084X DO - 10.7554/eLife.18164 UR - https://m2.mtmt.hu/api/publication/3083629 ID - 3083629 AB - In CFTR, the chloride channel mutated in cystic fibrosis (CF) patients, ATP-binding-induced dimerization of two cytosolic nucleotide binding domains (NBDs) opens the pore, and dimer disruption following ATP hydrolysis closes it. Spontaneous openings without ATP are rare in wild-type CFTR, but in certain CF mutants constitute the only gating mechanism, stimulated by ivacaftor, a clinically approved CFTR potentiator. The molecular motions underlying spontaneous gating are unclear. Here we correlate energetic coupling between residues across the dimer interface with spontaneous pore opening/closure in single CFTR channels. We show that spontaneous openings are also strictly coupled to NBD dimerization, which may therefore occur even without ATP. Coordinated NBD/pore movements are therefore intrinsic to CFTR: ATP alters the stability, but not the fundamental structural architecture, of open- and closed-pore conformations. This explains correlated effects of phosphorylation, mutations, and drugs on ATP-driven and spontaneous activity, providing insights for understanding CF mutation and drug mechanisms. LA - English DB - MTMT ER - TY - JOUR AU - Szabó, Eszter AU - Mizsei, Réka AU - Zámbó, Zsófia Melinda AU - Törőcsik, Beáta AU - Weiss, Manfred S AU - Ádám, Veronika AU - Ambrus, Attila TI - Crystal structure of the D444V disease-causing mutant of human dihydrolipoamide dehydrogenase JF - BIOCHIMICA ET BIOPHYSICA ACTA-BIOENERGETICS J2 - BBA-BIOENERGETICS VL - 1857 PY - 2016 IS - Suppl. SP - e100 SN - 0005-2728 DO - 10.1016/j.bbabio.2016.04.337 UR - https://m2.mtmt.hu/api/publication/3262509 ID - 3262509 LA - English DB - MTMT ER - TY - JOUR AU - Ambrus, Attila AU - Nemeria, NS AU - Törőcsik, Beáta AU - Tretter, László AU - Nilsson, M AU - Jordan, F AU - Ádám, Veronika TI - Formation of reactive oxygen species by human and bacterial pyruvate and 2- oxoglutarate dehydrogenase multienzyme complexes reconstituted from recombinant components JF - FREE RADICAL BIOLOGY AND MEDICINE J2 - FREE RADICAL BIO MED VL - 89 PY - 2015 SP - 642 EP - 650 PG - 9 SN - 0891-5849 DO - 10.1016/j.freeradbiomed.2015.10.001 UR - https://m2.mtmt.hu/api/publication/2953634 ID - 2953634 N1 - 02001, MTA, Magyar Tudományos Akadémia; 112230, OTKA, Magyar Tudományos Akadémia Department of Medical Biochemistry, MTA-SE Laboratory for Neurobiochemistry, Semmelweis University, Budapest, 1094, Hungary Department of Chemistry, Rutgers, State University, Newark, NJ 07102, United States Cited By :25 Export Date: 28 May 2020 CODEN: FRBME Correspondence Address: Adam-Vizi, V.; Department of Chemistry, Rutgers, State UniversityUnited States; email: adam.veronika@med.semmelweis-univ.hu Chemicals/CAS: adenosine diphosphate, 20398-34-9, 58-64-0; calcium ion, 14127-61-8; cytochrome c, 9007-43-6, 9064-84-0; oxoglutarate dehydrogenase, 9031-02-1; pyruvic acid, 127-17-3, 19071-34-2, 57-60-3; reduced nicotinamide adenine dinucleotide, 58-68-4; superoxide, 11062-77-4; Ketoglutarate Dehydrogenase Complex; Multienzyme Complexes; Pyruvic Acid; Reactive Oxygen Species; Recombinant Proteins Funding details: Magyar Tudományos Akadémia, MTA Funding details: Hungarian Scientific Research Fund, OTKA Funding details: Hungarian Scientific Research Fund, OTKA, 112230 Funding details: Magyar Tudományos Akadémia, MTA, 02001 Funding details: KTIA_13_NAP-A-III/6 Funding text 1: We are grateful to Drs. Hetalben Patel, Sowmini Kumaran, Junjie Wang, all from Rutgers, and Da Jeong Shim and Edgardo T. Farinas (New Jersey Institute of Technology) for providing purified proteins to this project, and Mulchand S. Patel and his group for providing cells harboring plasmids of the hPDHc components. Financial support is gratefully acknowledged from the Hungarian Academy of Sciences (MTA grant 02001 to A-V.V.), the Hungarian Scientific Research Fund (OTKA, grant 112230 to A-V. V.), the Hungarian Brain Research Program (grant KTIA_13_NAP-A-III/6. to A-V.V.), the Bolyai and the Fulbright Fellowships (to A.A.), and the NIH (NIH-GM-050380 and NIH-GM-116077 to F.J.). Department of Medical Biochemistry, MTA-SE Laboratory for Neurobiochemistry, Semmelweis University, Budapest, 1094, Hungary Department of Chemistry, Rutgers, State University, Newark, NJ 07102, United States Cited By :26 Export Date: 5 January 2021 CODEN: FRBME Correspondence Address: Adam-Vizi, V.; Department of Chemistry, Rutgers, State UniversityUnited States; email: adam.veronika@med.semmelweis-univ.hu Chemicals/CAS: adenosine diphosphate, 20398-34-9, 58-64-0; calcium ion, 14127-61-8; cytochrome c, 9007-43-6, 9064-84-0; oxoglutarate dehydrogenase, 9031-02-1; pyruvic acid, 127-17-3, 19071-34-2, 57-60-3; reduced nicotinamide adenine dinucleotide, 58-68-4; superoxide, 11062-77-4; Ketoglutarate Dehydrogenase Complex; Multienzyme Complexes; Pyruvic Acid; Reactive Oxygen Species; Recombinant Proteins Funding details: Magyar Tudományos Akadémia, MTA Funding details: Hungarian Scientific Research Fund, OTKA Funding details: Hungarian Scientific Research Fund, OTKA, 112230 Funding details: Magyar Tudományos Akadémia, MTA, 02001 Funding details: KTIA_13_NAP-A-III/6 Funding text 1: We are grateful to Drs. Hetalben Patel, Sowmini Kumaran, Junjie Wang, all from Rutgers, and Da Jeong Shim and Edgardo T. Farinas (New Jersey Institute of Technology) for providing purified proteins to this project, and Mulchand S. Patel and his group for providing cells harboring plasmids of the hPDHc components. Financial support is gratefully acknowledged from the Hungarian Academy of Sciences (MTA grant 02001 to A-V.V.), the Hungarian Scientific Research Fund (OTKA, grant 112230 to A-V. V.), the Hungarian Brain Research Program (grant KTIA_13_NAP-A-III/6. to A-V.V.), the Bolyai and the Fulbright Fellowships (to A.A.), and the NIH (NIH-GM-050380 and NIH-GM-116077 to F.J.). LA - English DB - MTMT ER - TY - JOUR AU - Csanády, László AU - Törőcsik, Beáta TI - Structure-activity analysis of a CFTR channel potentiator: Distinct molecular parts underlie dual gating effects. JF - JOURNAL OF GENERAL PHYSIOLOGY J2 - J GEN PHYSIOL VL - 144 PY - 2014 IS - 4 SP - 321 EP - 336 PG - 16 SN - 0022-1295 DO - 10.1085/jgp.201411246 UR - https://m2.mtmt.hu/api/publication/2761953 ID - 2761953 AB - The cystic fibrosis (CF) transmembrane conductance regulator (CFTR) is a member of the ATP-binding cassette transporter superfamily that functions as an epithelial chloride channel. Gating of the CFTR ion conduction pore involves a conserved irreversible cyclic mechanism driven by ATP binding and hydrolysis at two cytosolic nucleotide-binding domains (NBDs): formation of an intramolecular NBD dimer that occludes two ATP molecules opens the pore, whereas dimer disruption after ATP hydrolysis closes it. CFTR dysfunction resulting from inherited mutations causes CF. The most common CF mutation, deletion of phenylalanine 508 (DeltaF508), impairs both protein folding and processing and channel gating. Development of DeltaF508 CFTR correctors (to increase cell surface expression) and potentiators (to enhance open probability, Po) is therefore a key focus of CF research. The practical utility of 5-nitro-2-(3-phenylpropylamino)benzoate (NPPB), one of the most efficacious potentiators of DeltaF508 CFTR identified to date, is limited by its pore-blocking side effect. NPPB-mediated stimulation of Po is unique in that it involves modulation of gating transition state stability. Although stabilization by NPPB of the transition state for pore opening enhances both the rate of channel opening and the very slow rate of nonhydrolytic closure, because of CFTR's cyclic gating mechanism, the net effect is Po stimulation. In addition, slowing of ATP hydrolysis by NPPB delays pore closure, further enhancing Po. Here we show that NPPB stimulates gating at a site outside the pore and that these individual actions of NPPB on CFTR are fully attributable to one or the other of its two complementary molecular parts, 3-nitrobenzoate (3NB) and 3-phenylpropylamine (3PP), both of which stimulate Po: the pore-blocking 3NB selectively stabilizes the transition state for opening, whereas the nonblocking 3PP selectively slows the ATP hydrolysis step. Understanding structure-activity relationships of NPPB might prove useful for designing potent, clinically relevant CFTR potentiators. LA - English DB - MTMT ER - TY - JOUR AU - Csanády, László AU - Törőcsik, Beáta TI - Catalyst-like Modulation of Transitions States for CFTR Channel Opening and Closing: New Stimulation Strategy Exploits Nonequilibrium Gating JF - JOURNAL OF GENERAL PHYSIOLOGY J2 - J GEN PHYSIOL VL - 143 PY - 2014 IS - 2 SP - 269 EP - 287 PG - 19 SN - 0022-1295 DO - 10.1085/jgp.201311089 UR - https://m2.mtmt.hu/api/publication/2496247 ID - 2496247 LA - English DB - MTMT ER - TY - JOUR AU - Csanády, László AU - Mihályi, Csaba AU - Szöllősi, András AU - Törőcsik, Beáta AU - Vergani, P TI - Conformational changes in the catalytically inactive nucleotide-binding site of CFTR JF - JOURNAL OF GENERAL PHYSIOLOGY J2 - J GEN PHYSIOL VL - 142 PY - 2013 IS - 1 SP - 61 EP - 73 PG - 13 SN - 0022-1295 DO - 10.1085/jgp.201210954 UR - https://m2.mtmt.hu/api/publication/2333381 ID - 2333381 AB - A central step in the gating of the cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel is the association of its two cytosolic nucleotide-binding domains (NBDs) into a head-to-tail dimer, with two nucleotides bound at the interface. Channel opening and closing, respectively, are coupled to formation and disruption of this tight NBD dimer. CFTR is an asymmetric adenosine triphosphate (ATP)-binding cassette protein in which the two interfacial-binding sites (composite sites 1 and 2) are functionally different. During gating, the canonical, catalytically active nucleotide-binding site (site 2) cycles between dimerized prehydrolytic (state O1), dimerized post-hydrolytic (state O2), and dissociated (state C) forms in a preferential C-->O1-->O2-->C sequence. In contrast, the catalytically inactive nucleotide-binding site (site 1) is believed to remain associated, ATP-bound, for several gating cycles. Here, we have examined the possibility of conformational changes in site 1 during gating, by studying gating effects of perturbations in site 1.Previous work showed that channel closure is slowed, both under hydrolytic and nonhydrolytic conditions, by occupancy of site 1 by N6-(2-phenylethyl)-ATP (P-ATP) as well as by the site-1 mutation H1348A (NBD2 signature sequence). Here, we found that P-ATP prolongs wild-type (WT) CFTR burst durations by selectively slowing (>2x) transition O1-->O2 and decreases the nonhydrolytic closing rate (transition O1-->C) of CFTR mutants K1250A ( approximately 4x) and E1371S ( approximately 3x). Mutation H1348A also slowed ( approximately 3x) the O1-->O2 transition in the WT background and decreased the nonhydrolytic closing rate of both K1250A ( approximately 3x) and E1371S ( approximately 3x) background mutants. Neither P-ATP nor the H1348A mutation affected the 1:1 stoichiometry between ATP occlusion and channel burst events characteristic to WT CFTR gating in ATP. The marked effect that different structural perturbations at site 1 have on both steps O1-->C and O1-->O2 suggests that the overall conformational changes that CFTR undergoes upon opening and coincident with hydrolysis at the active site 2 include significant structural rearrangement at site 1. LA - English DB - MTMT ER - TY - JOUR AU - Kucharczyk, Roza AU - Wysocka-Kapcinska, M AU - Törőcsik, Beáta AU - Turiák, Lilla AU - Tsaprailis, G AU - David, C L AU - Hunt, A AU - Vékey, Károly AU - Ádám, Veronika AU - Chinopoulos, Christos TI - SAL1 modulates sensitivity of heterologously expressed artemia ADP/ATP carrier to bongkrekate in yeast JF - YEAST J2 - YEAST VL - 30 PY - 2013 IS - Suppl 1 SP - 164 EP - 164 PG - 1 SN - 0749-503X UR - https://m2.mtmt.hu/api/publication/2528949 ID - 2528949 LA - English DB - MTMT ER - TY - JOUR AU - Wysocka-Kapcinska, M AU - Törőcsik, Beáta AU - Turiák, Lilla AU - Tsaprailis, G AU - David, CL AU - Hunt, AM AU - Vékey, Károly AU - Ádám, Veronika AU - Kucharczyk, R AU - Chinopoulos, Christos TI - The Suppressor of AAC2 Lethality SAL1 Modulates Sensitivity of Heterologously Expressed Artemia ADP/ATP Carrier to Bongkrekate in Yeast JF - PLOS ONE J2 - PLOS ONE VL - 8 PY - 2013 IS - 9 PG - 12 SN - 1932-6203 DO - 10.1371/journal.pone.0074187 UR - https://m2.mtmt.hu/api/publication/2420731 ID - 2420731 N1 - Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland Department of Medical Biochemistry, Semmelweis University, Budapest, Hungary Institute of Organic Chemistry, Research Centre for Natural Sciences, Hungarian Academy of Sciences, Budapest, Hungary University of Arizona, Center for Toxicology, College of Pharmacy, Tucson, AZ, United States Cited By :5 Export Date: 17 December 2019 CODEN: POLNC Correspondence Address: Kucharczyk, R.; Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland; email: roza@ibb.waw.pl Chemicals/CAS: adenine nucleotide translocase, 9068-80-8; bongkrekic acid, 11076-19-0; hemagglutinin, 37333-12-3 Funding details: National Institute of Environmental Health Sciences, NIEHS, ES06694 AB - The ADP/ATP carrier protein (AAC) expressed in Artemia franciscana is refractory to bongkrekate. We generated two strains of Saccharomyces cerevisiae where AAC1 and AAC3 were inactivated and the AAC2 isoform was replaced with Artemia AAC containing a hemagglutinin tag (ArAAC-HA). In one of the strains the suppressor of ΔAAC2 lethality, SAL1, was also inactivated but a plasmid coding for yeast AAC2 was included, because the ArAACΔsal1Δ strain was lethal. In both strains ArAAC-HA was expressed and correctly localized to the mitochondria. Peptide sequencing of ArAAC expressed in Artemia and that expressed in the modified yeasts revealed identical amino acid sequences. The isolated mitochondria from both modified strains developed 85% of the membrane potential attained by mitochondria of control strains, and addition of ADP yielded bongkrekate-sensitive depolarizations implying acquired sensitivity of ArAAC-mediated adenine nucleotide exchange to this poison, independent from SAL1. However, growth of ArAAC-expressing yeasts in glycerol-containing media was arrested by bongkrekate only in the presence of SAL1. We conclude that the mitochondrial environment of yeasts relying on respiratory growth conferred sensitivity of ArAAC to bongkrekate in a SAL1-dependent manner. © 2013 Wysocka-Kapcinska et al. LA - English DB - MTMT ER - TY - JOUR AU - Konrád, Csaba AU - Kiss, Gergely AU - Törőcsik, Beáta AU - Ádám, Veronika AU - Chinopoulos, Christos TI - Absence of Ca2+-induced mitochondrial permeability transition but presence of bongkrekate-sensitive nucleotide exchange in C. crangon and P. serratus JF - BIOCHIMICA ET BIOPHYSICA ACTA-BIOENERGETICS J2 - BBA-BIOENERGETICS VL - 1817 PY - 2012 IS - Suppl. SP - S121 EP - S121 SN - 0005-2728 DO - 10.1016/j.bbabio.2012.06.325 UR - https://m2.mtmt.hu/api/publication/2529232 ID - 2529232 N1 - 17th European Bioenergetics Conference SEP 15-20, 2012 Freiburg, GERMANY LA - English DB - MTMT ER - TY - JOUR AU - Konrád, Csaba AU - Kiss, Gergely AU - Törőcsik, Beáta AU - Ádám, Veronika AU - Chinopoulos, Christos TI - Absence of Ca2+-induced mitochondrial permeability transition but presence of bongkrekate-sensitive nucleotide exchange in C. crangon and P. serratus JF - PLOS ONE J2 - PLOS ONE VL - 7 PY - 2012 IS - 6 PG - 9 SN - 1932-6203 DO - 10.1371/journal.pone.0039839 UR - https://m2.mtmt.hu/api/publication/1980192 ID - 1980192 LA - English DB - MTMT ER - TY - JOUR AU - Ambrus, Attila AU - Törőcsik, Beáta AU - Tretter, László AU - Ozohanics, Olivér AU - Ádám, Veronika TI - Stimulation of reactive oxygen species generation by disease-causing mutations of lipoamide dehydrogenase JF - HUMAN MOLECULAR GENETICS J2 - HUM MOL GENET VL - 20 PY - 2011 IS - 15 SP - 2984 EP - 2995 PG - 12 SN - 0964-6906 DO - 10.1093/hmg/ddr202 UR - https://m2.mtmt.hu/api/publication/1666747 ID - 1666747 LA - English DB - MTMT ER - TY - JOUR AU - Chinopoulos, Christos AU - Konrád, Csaba AU - Kiss, Gergely AU - Metelkin, E AU - Törőcsik, Beáta AU - Zhang, SF AU - Starkov, AA TI - Modulation of F0F1-ATP synthase activity by cyclophilin D regulates matrix adenine nucleotide levels JF - FEBS JOURNAL J2 - FEBS J VL - 278 PY - 2011 IS - 7 SP - 1112 EP - 1125 PG - 14 SN - 1742-464X DO - 10.1111/j.1742-4658.2011.08026.x UR - https://m2.mtmt.hu/api/publication/1502467 ID - 1502467 LA - English DB - MTMT ER - TY - JOUR AU - Dóczi, Judit AU - Turiák, Lilla AU - Sisa-Vajda, Szilvia AU - Mándi, Miklós AU - Törőcsik, Beáta AU - Gerencsér, Ákos AU - Kiss, Gergely AU - Konrád, Csaba AU - Ádám, Veronika AU - Chinopoulos, Christos TI - Complex Contribution of Cyclophilin D to Ca2+-induced Permeability Transition in Brain Mitochondria, with Relation to the Bioenergetic State JF - JOURNAL OF BIOLOGICAL CHEMISTRY J2 - J BIOL CHEM VL - 286 PY - 2011 IS - 8 SP - 6345 EP - 6353 PG - 9 SN - 0021-9258 DO - 10.1074/jbc.M110.196600 UR - https://m2.mtmt.hu/api/publication/1479724 ID - 1479724 LA - English DB - MTMT ER - TY - JOUR AU - Konrád, Csaba AU - Kiss, Gergely AU - Törőcsik, Beáta AU - Lábár, János AU - Gerencsér, Ákos AU - Mándi, Miklós AU - Ádám, Veronika AU - Chinopoulos, Christos TI - A distinct sequence in the adenine nucleotide translocase from Artemia franciscana embryos is associated with insensitivity to bongkrekate and atypical effects of adenine nucleotides on Ca2+ uptake and sequestration JF - FEBS JOURNAL J2 - FEBS J VL - 278 PY - 2011 IS - 5 SP - 822 EP - 836 PG - 15 SN - 1742-464X DO - 10.1111/j.1742-4658.2010.08001.x UR - https://m2.mtmt.hu/api/publication/1479725 ID - 1479725 LA - English DB - MTMT ER - TY - JOUR AU - Chinopoulos, Christos AU - Gerencsér, Ákos AU - Mándi, Miklós AU - Mathe, K AU - Törőcsik, Beáta AU - Dóczi, Judit AU - Turiák, Lilla AU - Kiss, Gergely AU - Konrád, Csaba AU - Sisa-Vajda, Szilvia AU - Vereczki, Viktória AU - Oh, RJ AU - Ádám, Veronika TI - Forward operation of adenine nucleotide translocase during F0F1-ATPase reversal: critical role of matrix substrate-level phosphorylation JF - FASEB JOURNAL J2 - FASEB J VL - 24 PY - 2010 IS - 7 SP - 2405 EP - 2416 PG - 12 SN - 0892-6638 DO - 10.1096/fj.09-149898 UR - https://m2.mtmt.hu/api/publication/1327780 ID - 1327780 LA - English DB - MTMT ER - TY - JOUR AU - Konrád, Csaba AU - Kiss, Gergely AU - Törőcsik, Beáta AU - Lábár, János AU - Gerencser, AA AU - Mándi, Miklós AU - Ádám, Veronika AU - Chinopoulos, Christos TI - Mitochondria from Artemia franciscana embryos exhibit a truncated form of ant, associated with atypical effects of its ligands on Ca2+ uptake capacity and unique morphology of matrix Ca2+ precipitates JF - BIOCHIMICA ET BIOPHYSICA ACTA-BIOENERGETICS J2 - BBA-BIOENERGETICS VL - 1797 PY - 2010 SP - 142 EP - 143 PG - 2 SN - 0005-2728 DO - 10.1016/j.bbabio.2010.04.426 UR - https://m2.mtmt.hu/api/publication/1404329 ID - 1404329 LA - English DB - MTMT ER - TY - JOUR AU - Ambrus, Attila AU - Törőcsik, Beáta AU - Ádám, Veronika TI - Refolding of the human dihydrolipoamide dehydrogenase JF - BIOCHEMICAL ENGINEERING JOURNAL J2 - BIOCHEM ENG J VL - 45 PY - 2009 IS - 2 SP - 120 EP - 125 PG - 6 SN - 1369-703X DO - 10.1016/j.bej.2009.03.004 UR - https://m2.mtmt.hu/api/publication/1264449 ID - 1264449 LA - English DB - MTMT ER - TY - JOUR AU - Ambrus, Attila AU - Törőcsik, Beáta AU - Ádám, Veronika TI - Periplasmic cold expression and one-step purification of human dihydrolipoamide dehydrogenase JF - PROTEIN EXPRESSION AND PURIFICATION J2 - PROTEIN EXPRES PURIF VL - 63 PY - 2009 IS - 1 SP - 50 EP - 57 PG - 8 SN - 1046-5928 DO - 10.1016/j.pep.2008.09.009 UR - https://m2.mtmt.hu/api/publication/244260 ID - 244260 AB - Dihydrolipoamide dehydrogenase (LADH) is a FAD-linked subunit of alpha-ketoglutarate, pyruvate and branched-chain amino acid dehydrogenases and the glycine cleavage system. As an oxidoreductase it transfers electrons from the dihydrolipoic acid prosthetic group to the NAD(+) cofactor via its FAD center. Besides its physiological function it is capable of generating harmful reactive oxygen species (ROS) in pathological settings therefore it is implicated in neurodegeneration, ischemia- reperfusion, cancer and several other disorders. Pathological mutants of the enzyme cause severe, sometimes lethal syndromes like hypotonia, metabolic acidosis or inefficiency in development. Recently it has been revealed that LADH is a moonlighting protease when specific mutations in the dimerization surface destabilize the functional homodimer and expose a serine-protease-like catalytic dyad. As the basis of versatile functions of LADH is far from elucidation, there is a constant need for a pure and functional enzyme product for investigations. Several studies used recombinant human LADH before, however, it was generated by more complicated and/or physiologically less compatible protocols than reported here; most papers on functional and structural studies do not even report detailed protocols and characteristics (most importantly the purity) of their protein products. Here we describe the details of an optimized, easy-to-use periplasmic expression and one-step purification protocol for obtaining a highly pure, active and authentic (tag-cleaved) enzyme with the characterization of the protein product. The purified LADH can be used in biophysical and structural studies while the published protocol is easily convertible to a protein labeling procedure. (c) 2008 Elsevier Inc. All rights reserved. LA - English DB - MTMT ER - TY - JOUR AU - Csanády, László AU - Törőcsik, Beáta TI - Four Ca2+ Ions Activate TRPM2 Channels by Binding in Deep Crevices near the Pore but Intracellularly of the Gate JF - JOURNAL OF GENERAL PHYSIOLOGY J2 - J GEN PHYSIOL VL - 133 PY - 2009 IS - 2 SP - 189 EP - 203 PG - 15 SN - 0022-1295 DO - 10.1085/jgp.200810109 UR - https://m2.mtmt.hu/api/publication/1502468 ID - 1502468 LA - English DB - MTMT ER - TY - JOUR AU - Gerencsér, Ákos AU - Dóczi, Judit AU - Törőcsik, Beáta AU - Bossy, B AU - Bossy-Wetzel, E AU - Ádám, Veronika TI - Mitochondrial swelling measurement in situ by optimized spatial filtering: Astrocyte-neuron differences JF - BIOPHYSICAL JOURNAL J2 - BIOPHYS J VL - 95 PY - 2008 IS - 5 SP - 2583 EP - 2598 PG - 16 SN - 0006-3495 DO - 10.1529/biophysj.107.118620 UR - https://m2.mtmt.hu/api/publication/1131095 ID - 1131095 LA - English DB - MTMT ER - TY - JOUR AU - Feketéné Kiss, Katalin AU - Salamon, S AU - Törőcsik, Beáta AU - Szeberényi, József TI - Role of phospholipase C-gamma in NGF-stimulated differentiation and gene induction JF - ACTA BIOLOGICA HUNGARICA (1983-2018) J2 - ACTA BIOL HUNG VL - 57 PY - 2006 IS - 2 SP - 147 EP - 155 PG - 9 SN - 0236-5383 DO - 10.1556/ABiol.57.2006.2.2 UR - https://m2.mtmt.hu/api/publication/1428260 ID - 1428260 AB - The PC12 phaeochromocytoma cell line provides a useful model to study nerve growth factor-induced neuronal differentiation. The central signaling route of this process is mediated by the Ras-dependent extracellular signal-regulated kinase cascade. However, Ras-independent pathways are also stimulated by nerve growth factor and may contribute to differentiation signaling. One mediator for Ras-independent signal transduction in PC12 cells is phospholipase C-gamma that generates the second messengers diacylglycerol and inositol- trisphosphate. To probe the possible involvement of this enzyme in nerve growth factor-promoted differentiation, we used the phospholipase C inhibitor U73122 and the inositol-trisphosphatereceptor inhibitor Xestospongin C. Our results show that both chemicals block nerve growth factor-promoted neurite out-growth, but the blockage of phospholipase C does not inhibit nerve growth factor-induced expression of c-fos, zif268 and transin genes. In addition, induction of these genes by nerve growth factor plus dibutyryl-cAMP is comparable in wild-type PC12 cells as well as in cells in which both Ras- and phospholipase C-gamma-mediated pathways are inhibited. The phospholipase C-gamma pathway thus belongs to those nerve growth factor receptor-originated signaling routes that contribute to the biological response of PC12 cells to nerve growth factor, but its gene activating potential does not have a major role in its neuritogenic effect. LA - English DB - MTMT ER - TY - JOUR AU - Sipos, Ildikó AU - Törőcsik, Beáta AU - Tretter, László AU - Ádám, Veronika TI - Impaired regulation of pH homeostasis by oxidative stress in rat brain capillary endothelial cells JF - CELLULAR AND MOLECULAR NEUROBIOLOGY J2 - CELL MOL NEUROBIOL VL - 25 PY - 2005 IS - 1 SP - 141 EP - 151 PG - 11 SN - 0272-4340 DO - 10.1007/s10571-004-1379-6 UR - https://m2.mtmt.hu/api/publication/1030894 ID - 1030894 LA - English DB - MTMT ER - TY - JOUR AU - Szeberényi, József AU - Fábián, Zsolt AU - Törőcsik, Beáta AU - Feketéné Kiss, Katalin AU - Csatáry, L. K. TI - Newcastle disease virus-induced apoptosis in PC12 pheochromocytoma cells JF - AMERICAN JOURNAL OF THERAPEUTICS J2 - AM J THER VL - 10 PY - 2003 IS - 4 SP - 282 EP - 288 PG - 7 SN - 1075-2765 DO - 10.1097/00045391-200307000-00008 UR - https://m2.mtmt.hu/api/publication/1586978 ID - 1586978 LA - English DB - MTMT ER - TY - JOUR AU - Angelastro, JM AU - Ryu, EJ AU - Törőcsik, Beáta AU - Fiske, BK AU - Greene, LA TI - Blue-white selection step enhances the yield of SAGE concatemers JF - BIOTECHNIQUES J2 - BIOTECHNIQUES VL - 32 PY - 2002 IS - 3 SP - 484, 486 PG - 2 SN - 0736-6205 DO - 10.2144/02323bm02 UR - https://m2.mtmt.hu/api/publication/1502471 ID - 1502471 LA - English DB - MTMT ER - TY - JOUR AU - Angelastro, JM AU - Törőcsik, Beáta AU - Greene, LA TI - Nerve growth factor selectively regulates expression of transcripts encoding ribosomal proteins JF - BMC NEUROSCIENCE J2 - BMC NEUROSCI VL - 3 PY - 2002 PG - 11 SN - 1471-2202 DO - 10.1186/1471-2202-3-3 UR - https://m2.mtmt.hu/api/publication/1502469 ID - 1502469 LA - English DB - MTMT ER - TY - JOUR AU - Törőcsik, Beáta AU - Angelastro, JM AU - Greene, LA TI - The basic region and leucine zipper transcription factor MafK is a new nerve growth factor-responsive immediate early gene that regulates neurite outgrowth JF - JOURNAL OF NEUROSCIENCE J2 - J NEUROSCI VL - 22 PY - 2002 IS - 20 SP - 8971 EP - 8980 PG - 10 SN - 0270-6474 UR - https://m2.mtmt.hu/api/publication/1502470 ID - 1502470 LA - English DB - MTMT ER - TY - JOUR AU - Fábián, Zsolt AU - Törőcsik, Beáta AU - Feketéné Kiss, Katalin AU - Csatary, LK AU - Bodey, B AU - Tigyi, József AU - Csatary, C AU - Szeberényi, József TI - Induction of apoptosis by a Newcastle disease virus vaccine (MTH-68/H) in PC12 rat phaeochromocytoma cells JF - ANTICANCER RESEARCH J2 - ANTICANCER RES VL - 21 PY - 2001 IS - 1A SP - 125 EP - 135 PG - 11 SN - 0250-7005 UR - https://m2.mtmt.hu/api/publication/1502472 ID - 1502472 LA - English DB - MTMT ER - TY - JOUR AU - Oszter, Angéla AU - Törőcsik, Beáta AU - Vértes, Zsuzsanna AU - Környei, József László AU - Kovács, Kálmán András AU - Vértes, Marietta TI - Regulation of activator protein-1-DNA binding activity by opioid peptides in estrogen-sensitive cells of rat hypothalamus and uterus JF - EUROPEAN JOURNAL OF PHARMACOLOGY J2 - EUR J PHARMACOL VL - 395 PY - 2000 IS - 2 SP - 103 EP - 106 PG - 4 SN - 0014-2999 DO - 10.1016/S0014-2999(00)00214-4 UR - https://m2.mtmt.hu/api/publication/1427998 ID - 1427998 LA - English DB - MTMT ER - TY - JOUR AU - Oszter, Angéla AU - Vértes, Zsuzsanna AU - Törőcsik, Beáta AU - Környei, József László AU - Kovács, Kálmán András AU - Vértes, Marietta TI - Antiestrogenic effect of opioid peptides in rat uterus. JF - JOURNAL OF STEROID BIOCHEMISTRY AND MOLECULAR BIOLOGY J2 - J STEROID BIOCHEM MOL BIOL VL - 74 PY - 2000 IS - 1-2 SP - 25 EP - 32 PG - 8 SN - 0960-0760 DO - 10.1016/S0960-0760(00)00085-6 UR - https://m2.mtmt.hu/api/publication/1427993 ID - 1427993 LA - English DB - MTMT ER - TY - JOUR AU - Törőcsik, Beáta AU - Szeberényi, József TI - Anisomycin uses multiple mechanisms to stimulate mitogen-activated protein kinases and gene expression and to inhibit neuronal differentiation in PC12 phaeochromocytoma cells JF - EUROPEAN JOURNAL OF NEUROSCIENCE J2 - EUR J NEUROSCI VL - 12 PY - 2000 IS - 2 SP - 527 EP - 532 PG - 6 SN - 0953-816X DO - 10.1046/j.1460-9568.2000.00933.x UR - https://m2.mtmt.hu/api/publication/1428299 ID - 1428299 AB - Treatment of PC12 cells with nerve growth factor (NGF) stimulates extracellular signal-regulated kinases (ERKs), as well as stress-activated c-Jun N-terminal kinases (JNKs) and p38 kinase, and induces neuronal differentiation. While the pivotal role of ERKs in NGF-induced morphological differentiation is well established, the contribution of JNK- and p38-pathways is less clear. The role of the JNK- and p38-pathway in PC12 cells was analysed by using anisomycin, a protein synthesis inhibitor that activates JNKs and p38. Non-toxic concentrations of anisomycin were found to stimulate these enzyme activities as well as the expression of the early response genes c-jun, c-fos and zif268, and to inhibit NGF-induced neurite formation. These effects of anisomycin appear to be mediated by the generation of reactive oxygen species (ROS), which in turn act through both TrkA/Ras-dependent and -independent signalling pathways. In addition, cross-talk between the p38- and ERK-pathways appears to play a role in the action of anisomycin. LA - English DB - MTMT ER - TY - JOUR AU - Törőcsik, Beáta AU - Szeberényi, József TI - Anisomycin affects both pro- and antiapoptotic mechanisms in PC12 cells JF - BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS J2 - BIOCHEM BIOPH RES CO VL - 278 PY - 2000 IS - 3 SP - 550 EP - 556 PG - 7 SN - 0006-291X DO - 10.1006/bbrc.2000.3836 UR - https://m2.mtmt.hu/api/publication/1428298 ID - 1428298 AB - Survival and differentiation of PC12 cells depend on the proper balance between the activities of several mitogen-activated protein kinase (MAPK) pathways. We have previously shown that low, nontoxic doses of anisomycin stimulated these MAPKs as well. as the expression of several early-response genes and inhibited NC;F-induced neurite formation. In the present work we show that protein synthesis-inhibiting concentrations of anisomycin, in contrast, cause apoptosis of PC12 cells. To try to characterize the apoptosis-inducing mechanisms of anisomycin we compared the signaling effects of subinhibitory and inhibitory drug concentrations. Anisomycin in a nontoxic dosis activates the same MAPK pathways and early-response genes as in protein synthesis inhibiting concentrations. In contrast, while the subinhibitory anisomycin treatment stimulates Akt and induces Bcl-2, two anti-apoptotic proteins, the translation-inhibiting concentration of the drug prevents these survival-promoting biochemical events. Anisomycin thus triggers both pro- and anti-apoptotic processes in PC12 cells; stimulation of stress-responsive MAPK cascades is not sufficient to mediate apoptotic signaling: the inhibition of key antiapoptotic proteins appears to be more important for PC12 cell death by anisomycin treatment. (C) 2000 Academic Press. LA - English DB - MTMT ER - TY - JOUR AU - Csatary, LK AU - Moss, RW AU - Beuth, J AU - Törőcsik, Beáta AU - Szeberényi, József AU - Bakács, Tibor TI - Beneficial treatment of patients with advanced cancer using a Newcastle disease virus vaccine (MTH-68/H) JF - ANTICANCER RESEARCH J2 - ANTICANCER RES VL - 19 PY - 1999 IS - 1B SP - 635 EP - 638 PG - 4 SN - 0250-7005 UR - https://m2.mtmt.hu/api/publication/1339176 ID - 1339176 AB - Newcastle Disease Virus Vaccine (MTH-68/H) was administered to patients suffering from advanced neoplastic diseases after non-efficient tumor-destructive treatment. Case reports of selected patients suggest promising effects of this treatment. A prospectively-randomized clinical study (phase III; in accordance with Good Clinical Practice, GCP) was proposed to confirm these results and is currently under consideration. LA - English DB - MTMT ER -