TY - JOUR AU - Tóth, Ádám Viktor AU - Bartók, Ádám TI - Reviewing critical TRPM2 variants through a structure-function lens JF - JOURNAL OF BIOTECHNOLOGY J2 - J BIOTECHNOL VL - 385 PY - 2024 SP - 49 EP - 57 PG - 9 SN - 0168-1656 DO - 10.1016/j.jbiotec.2024.02.017 UR - https://m2.mtmt.hu/api/publication/34720489 ID - 34720489 LA - English DB - MTMT ER - TY - JOUR AU - Csanády, László TI - Towards personalized medicine for cystic fibrosis patients with rare mutations JF - JOURNAL OF PHYSIOLOGY-LONDON J2 - J PHYSIOL-LONDON VL - 602 PY - 2024 IS - 2 SP - 257 EP - 258 PG - 2 SN - 0022-3751 DO - 10.1113/JP286135 UR - https://m2.mtmt.hu/api/publication/34507016 ID - 34507016 LA - English DB - MTMT ER - TY - JOUR AU - Bartók, Ádám AU - Csanády, László TI - TRPM2 - an adjustable thermostat JF - CELL CALCIUM J2 - CELL CALCIUM VL - 118 PY - 2024 PG - 6 SN - 0143-4160 DO - 10.1016/j.ceca.2024.102850 UR - https://m2.mtmt.hu/api/publication/34502176 ID - 34502176 LA - English DB - MTMT ER - TY - JOUR AU - Simon, Márton AU - Iordanov, Iordan AU - Szöllősi, András AU - Csanády, László TI - Estimating the true stability of the prehydrolytic outward-facing state in an ABC protein. JF - ELIFE J2 - ELIFE VL - 12 PY - 2023 PG - 19 SN - 2050-084X DO - 10.7554/eLife.90736 UR - https://m2.mtmt.hu/api/publication/34232792 ID - 34232792 AB - CFTR, the anion channel mutated in cystic fibrosis patients, is a model ABC protein whose ATP-driven conformational cycle is observable at single-molecule level in patch-clamp recordings. Bursts of CFTR pore openings are coupled to tight dimerization of its two nucleotide-binding domains (NBDs) and in wild-type (WT) channels are mostly terminated by ATP hydrolysis. The slow rate of non-hydrolytic closure - which determines how tightly bursts and ATP hydrolysis are coupled - is unknown, as burst durations of catalytic site mutants span a range of ~200-fold. Here, we show that Walker A mutation K1250A, Walker B mutation D1370N, and catalytic glutamate mutations E1371S and E1371Q all completely disrupt ATP hydrolysis. True non-hydrolytic closing rate of WT CFTR approximates that of K1250A and E1371S. That rate is slowed ~15-fold in E1371Q by a non-native inter-NBD H-bond, and accelerated ~15-fold in D1370N. These findings uncover unique features of the NBD interface in human CFTR. LA - English DB - MTMT ER - TY - JOUR AU - Szöllősi, András AU - Almássy, János TI - Functional characterization of the transient receptor potential melastatin 2 (TRPM2) cation channel from Nematostella vectensis reconstituted into lipid bilayer JF - SCIENTIFIC REPORTS J2 - SCI REP VL - 13 PY - 2023 IS - 1 PG - 10 SN - 2045-2322 DO - 10.1038/s41598-023-38640-6 UR - https://m2.mtmt.hu/api/publication/34064647 ID - 34064647 N1 - Department of Biochemistry, Semmelweis University, Tuzolto u. 37-47, Budapest, 1094, Hungary Department of Physiology, Semmelweis University, Tuzolto u. 37-47, Budapest, 1094, Hungary ELKH-SE Ion Channel Research Group, Semmelweis University, Tuzolto u. 37-47, Budapest, 1094, Hungary HCEMM-SE Molecular Channelopathies Research Group, Semmelweis University, Tuzolto u. 37-47, Budapest, 1094, Hungary Export Date: 4 September 2023 Correspondence Address: Szollosi, A.; Department of Biochemistry, Tuzolto u. 37-47, Hungary; email: szollo5@gmail.com Chemicals/CAS: adenosine diphosphate ribose, 20762-30-5; calcium, 7440-70-2, 14092-94-5; Adenosine Diphosphate Ribose; Calcium; Cations; Lipid Bilayers; TRPM Cation Channels Funding details: 739593 Funding details: Nemzeti Kutatási, Fejlesztési és Innovaciós Alap, NKFIA, KKP 144199 Funding text 1: This work was supported by Semmelweis Science and Innovation Fund STIA_18_M; EU Horizon 2020 Research and Innovation Program grant 739593 and National Research, Development and Innovation Fund grant KKP 144199. We thank László Csanády for funding and thorough reading and comments on the manuscript. We thank Balázs Tóth for help with poly- l -lysine measurements and stimulating discussions. AB - Transient receptor potential melastatin 2 (TRPM2) cation channel activity is required for insulin secretion, immune cell activation and body heat control. Channel activation upon oxidative stress is involved in the pathology of stroke and neurodegenerative disorders. Cytosolic Ca 2+ , ADP-ribose (ADPR) and phosphatidylinositol-4,5-bisphosphate (PIP 2 ) are the obligate activators of the channel. Several TRPM2 cryo-EM structures have been resolved to date, yet functionality of the purified protein has not been tested. Here we reconstituted overexpressed and purified TRPM2 from Nematostella vectensis (nvTRPM2) into lipid bilayers and found that the protein is fully functional. Consistent with the observations in native membranes, nvTRPM2 in lipid bilayers is co-activated by cytosolic Ca 2+ and either ADPR or ADPR-2′-phosphate (ADPRP). The physiological metabolite ADPRP has a higher apparent affinity than ADPR. In lipid bilayers nvTRPM2 displays a large linear unitary conductance, its open probability (P o ) shows little voltage dependence and is stable over several minutes. P o is high without addition of exogenous PIP 2 , but is largely blunted by treatment with poly- l -Lysine, a polycation that masks PIP 2 headgroups. These results indicate that PIP 2 or some other activating phosphoinositol lipid co-purifies with nvTRPM2, suggesting a high PIP 2 binding affinity of nvTRPM2 under physiological conditions. LA - English DB - MTMT ER - TY - JOUR AU - Csanády, László TI - Blue flash sheds light on the roles of individual phosphoserines in CFTR channel activation JF - JOURNAL OF GENERAL PHYSIOLOGY J2 - J GEN PHYSIOL VL - 155 PY - 2023 IS - 6 PG - 3 SN - 0022-1295 DO - 10.1085/jgp.202313336 UR - https://m2.mtmt.hu/api/publication/33755807 ID - 33755807 N1 - Department of Biochemistry, Semmelweis University, Budapest, Hungary HCEMM-SE Molecular Channelopathies Research Group, Budapest, Hungary ELKH-SE Ion Channel Research Group, Budapest, Hungary Export Date: 31 August 2023 CODEN: JEMEA Correspondence Address: Csanády, L.; Department of Biochemistry, Hungary; email: csanady.laszlo@med.semmelweis-univ.hu Chemicals/CAS: cyclic AMP, 60-92-4; cystic fibrosis transmembrane conductance regulator, 126880-72-6; phosphoserine, 407-41-0; serine, 56-45-1, 6898-95-9; Cystic Fibrosis Transmembrane Conductance Regulator; Phosphoserine; Serine Funding details: 739593 Funding details: Cystic Fibrosis Foundation, CFF, CSANAD21G0 Funding details: Nemzeti Kutatási Fejlesztési és Innovációs Hivatal, NKFIH, 144199 Funding text 1: This work was supported by EU Horizon 2020 Research and Innovation Program grant 739593, Cystic Fibrosis Foundation Research Grant CSANAD21G0, and Nemzeti Kutatási Fejlesztési és Innovációs Hivatal KKP_22 grant 144199 to L. Csanády. The author declares no competing financial interests. LA - English DB - MTMT ER - TY - JOUR AU - Bartók, Ádám TI - Investigation of factors that influence the temperature sensitivity of the TRPM2 cation channel JF - BIOPHYSICAL JOURNAL J2 - BIOPHYS J VL - 122 PY - 2023 IS - 3S1 SP - 110a EP - 110a SN - 0006-3495 DO - 10.1016/j.bpj.2022.11.775 UR - https://m2.mtmt.hu/api/publication/33666263 ID - 33666263 N1 - Export Date: 27 February 2023 LA - English DB - MTMT ER - TY - JOUR AU - Simon, Márton AU - Csanády, László TI - Optimization of CFTR gating through the evolution of its extracellular loops JF - JOURNAL OF GENERAL PHYSIOLOGY J2 - J GEN PHYSIOL VL - 155 PY - 2023 IS - 4 PG - 16 SN - 0022-1295 DO - 10.1085/jgp.202213264 UR - https://m2.mtmt.hu/api/publication/33636409 ID - 33636409 LA - English DB - MTMT ER - TY - JOUR AU - Bartók, Ádám AU - Csanády, László TI - Dual amplification strategy turns TRPM2 channels into supersensitive central heat detectors JF - PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA J2 - P NATL ACAD SCI USA VL - 119 PY - 2022 IS - 48 PG - 11 SN - 0027-8424 DO - 10.1073/pnas.2212378119 UR - https://m2.mtmt.hu/api/publication/33262968 ID - 33262968 N1 - Hungarian Centre of Excellence for Molecular Medicine-Semmelweis Egyetem (HCEMM-SE), Molecular Channelopathies Research Group, Semmelweis University, Budapest, H-1094, Hungary Magyar Tudományos Akadémia-Semmelweis Egyetem (MTA-SE), Ion Channel Research Group, Semmelweis University, Budapest, H-1094, Hungary Department of Biochemistry, Semmelweis University, Budapest, H-1094, Hungary Cited By :3 Export Date: 4 September 2023 CODEN: PNASA Correspondence Address: Csanády, L.; Hungarian Centre of Excellence for Molecular Medicine-Semmelweis Egyetem (HCEMM-SE), Hungary; email: csanady.laszlo@med.semmelweis-univ.hu Chemicals/CAS: adenosine diphosphate ribose, 20762-30-5; calcium ion, 14127-61-8; hydrogen peroxide, 7722-84-1; calcium, 7440-70-2, 14092-94-5; Adenosine Diphosphate Ribose; Calcium; Hydrogen Peroxide; TRPM Cation Channels Funding details: 739593 Funding details: GINOP-2.3.2-15-2016-00051 Funding details: ÚNKP-20-5-SE-6, ÚNKP-21-5-SE-10 Funding details: Magyar Tudományos Akadémia, MTA, BO/00103/20, LP2017-14/2017 Funding details: Nemzeti Kutatási Fejlesztési és Innovációs Hivatal, NKFIH Funding text 1: We thank Beáta Töröcsik for subcloning T5L-TRPM2 into pcDNA3, Iordan Iordanov for providing purified nvNUDT9H, and Katalin Zboray (Agricultural Institute, Centre for Agricultural Research, Martonvásár, Hungary) for providing the stable T5L TRPM2 cell line (Economic Development and Innovation Operational Programme (GINOP-2.3.2-15-2016-00051) of the National Research, Development and Innovation Office). Support was provided by EU Horizon 2020 Research and Innovation Program grant 739593 and MTA Lendület grant LP2017-14/2017 to L.C. and a New National Excellence Program (ÚNKP) award of the Ministry of Human Capacities of Hungary to Semmelweis University. Á.B. was supported by the János Bolyai Research Scholarship of the Hungarian Academy of Sciences (BO/00103/20) and the New National Excellence Program (ÚNKP) Bolyai+ scholarship of the Ministry of Human Capacities of Hungary (ÚNKP-20-5-SE-6 and ÚNKP-21-5-SE-10). Funding text 2: ACKNOWLEDGMENTS. We thank Beáta Töröcsik for subcloning T5L-TRPM2 into pcDNA3, Iordan Iordanov for providing purified nvNUDT9H, and Katalin Zboray (Agricultural Institute, Centre for Agricultural Research, Martonvásár, Hungary) for providing the stableT5LTRPM2 cell line (Economic Development and Innovation Operational Programme (GINOP-2.3.2-15-2016-00051) of the National Research, Development and Innovation Office). Support was provided by EU Horizon 2020 Research and Innovation Program grant 739593 and MTA Lendület grant LP2017-14/2017 to L.C. and a New National Excellence Program (ÚNKP) award of the Ministry of Human Capacities of Hungary to Semmelweis University. Á.B. was supported by the János Bolyai Research Scholarship of the Hungarian Academy of Sciences (BO/00103/20) and the New National Excellence Program (ÚNKP) Bolyai+ scholarship of the Ministry of Human Capacities of Hungary (ÚNKP-20-5-SE-6 and ÚNKP-21-5-SE-10). AB - The Ca 2+ and ADP ribose (ADPR)-activated cation channel TRPM2 is the closest homolog of the cold sensor TRPM8 but serves as a deep-brain warmth sensor. To unravel the molecular mechanism of heat sensing by the TRPM2 protein, we study here temperature dependence of TRPM2 currents in cell-free membrane patches across ranges of agonist concentrations. We find that channel gating remains strictly agonist-dependent even at 40°C: heating alone or in combination with just Ca 2+ , just ADPR, Ca 2+ + cyclic ADPR, or H 2 O 2 pretreatment only marginally activates TRPM2. For fully liganded TRPM2, pore opening is intrinsically endothermic, due to ~10-fold larger activation enthalpy for opening (~200 kJ/mol) than for closure (~20 kJ/mol). However, the temperature threshold is too high (>40°C) for unliganded but too low (<15°C) for fully liganded channels. Thus, warmth sensitivity around 37°C is restricted to narrow ranges of agonist concentrations. For ADPR, that range matches, but for Ca 2+ , it exceeds bulk cytosolic values. The supraphysiological [Ca 2+ ] needed for TRPM2 warmth sensitivity is provided by Ca 2+ entering through the channel’s pore. That positive feedback provides further strong amplification to the TRPM2 temperature response (Q 10 ~ 1,000), enabling the TRPM2 protein to autonomously respond to tiny temperature fluctuations around 37°C. These functional data together with published structures suggest a molecular mechanism for opposite temperature dependences of two closely related channel proteins. LA - English DB - MTMT ER - TY - JOUR AU - Horváth, Gergő AU - Sváb, Gergely AU - Komlódi, Tímea AU - Ravasz, Dóra AU - Kacsó, Gergely AU - Dóczi, Judit AU - Chinopoulos, Christos AU - Ambrus, Attila AU - Tretter, László TI - Reverse and Forward Electron Flow-Induced H2O2 Formation Is Decreased in α-Ketoglutarate Dehydrogenase (α-KGDH) Subunit (E2 or E3) Heterozygote Knock Out Animals JF - ANTIOXIDANTS J2 - ANTIOXIDANTS-BASEL VL - 11 PY - 2022 IS - 8 PG - 19 SN - 2076-3921 DO - 10.3390/antiox11081487 UR - https://m2.mtmt.hu/api/publication/33070137 ID - 33070137 N1 - Cited By :1 Export Date: 7 October 2022 Correspondence Address: Tretter, L.; Department of Biochemistry, Hungary; email: tretter.laszlo@med.semmelweis-univ.hu Funding details: Semmelweis Egyetem, STIA-OTKA-2021, TKP2021-EGA-25 Funding details: Nemzeti Kutatási, Fejlesztési és Innovaciós Alap, NKFIA, EFOP-3.6.3-VEKOP-16-2017-00009 Funding text 1: This research was funded by the Hungarian Brain Research Program 2 (2017-1.2.1-NKP-2017-00002 to Vera Adam-Vizi, Semmelweis University), STIA-OTKA-2021 grant (from the Semmelweis University, to A.A.), TKP2021-EGA-25 grant to A.A. and C.C., 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. EFOP-3.6.3-VEKOP-16-2017-00009 support to G.S. AB - α-ketoglutarate dehydrogenase complex (KGDHc), or 2-oxoglutarate dehydrogenase complex (OGDHc) is a rate-limiting enzyme in the tricarboxylic acid cycle, that has been identified in neurodegenerative diseases such as in Alzheimer’s disease. The aim of the present study was to establish the role of the KGDHc and its subunits in the bioenergetics and reactive oxygen species (ROS) homeostasis of brain mitochondria. To study the bioenergetic profile of KGDHc, genetically modified mouse strains were used having a heterozygous knock out (KO) either in the dihydrolipoyl succinyltransferase (DLST+/−) or in the dihydrolipoyl dehydrogenase (DLD+/−) subunit. Mitochondrial oxygen consumption, hydrogen peroxide (H2O2) production, and expression of antioxidant enzymes were measured in isolated mouse brain mitochondria. Here, we demonstrate that the ADP-stimulated respiration of mitochondria was partially arrested in the transgenic animals when utilizing α-ketoglutarate (α-KG or 2-OG) as a fuel substrate. Succinate and α-glycerophosphate (α-GP), however, did not show this effect. The H2O2 production in mitochondria energized with α-KG was decreased after inhibiting the adenine nucleotide translocase and Complex I (CI) in the transgenic strains compared to the controls. Similarly, the reverse electron transfer (RET)-evoked H2O2 formation supported by succinate or α-GP were inhibited in mitochondria isolated from the transgenic animals. The decrease of RET-evoked ROS production by DLST+/− or DLD+/− KO-s puts the emphasis of the KGDHc in the pathomechanism of ischemia-reperfusion evoked oxidative stress. Supporting this notion, expression of the antioxidant enzyme glutathione peroxidase was also decreased in the KGDHc transgenic animals suggesting the attenuation of ROS-producing characteristics of KGDHc. These findings confirm the contribution of the KGDHc to the mitochondrial ROS production and in the pathomechanism of ischemia-reperfusion injury. LA - English DB - MTMT ER -