TY - JOUR AU - Lee, Derek C AU - Ta, Linh AU - Mukherjee, Purna AU - Duraj, Tomas AU - Domin, Marek AU - Greenwood, Bennett AU - Karmacharya, Srada AU - Narain, Niven R AU - Kiebish, Michael AU - Chinopoulos, Christos AU - Seyfried, Thomas N TI - Amino Acid and Glucose Fermentation Maintain ATP Content in Mouse and Human Malignant Glioma Cells. JF - ASN NEURO J2 - ASN NEURO VL - 16 PY - 2024 IS - 1 PG - 24 SN - 1759-0914 DO - 10.1080/17590914.2024.2422268 UR - https://m2.mtmt.hu/api/publication/35639179 ID - 35639179 N1 - Funding Agency and Grant Number: Foundation for Metabolic Cancer Therapies; Broken Science Initiative; Children with Cancer UK; Corkin Family Foundation; Boston College Research Expense Fund Funding text: We thank the Foundation for Metabolic Cancer Therapies, CrossFit Inc., Dr. Joseph C. Maroon, Dr. Edward Miller, The Broken Science Initiative, Children with Cancer UK, The Corkin Family Foundation, and the Boston College Research Expense Fund for their support. We also thank Miguel Estevez for providing the U-87MG cell line. We thank Dr. Laura Shelton for her original background work on the subject. We thank Bret Judson and the Boston College Imaging Core for their assistance with microscopy. Lastly, we thank the three anonymous reviewers for their input and suggestions, which improved the quality of our manuscript. AB - Energy is necessary for tumor cell viability and growth. Aerobic glucose-driven lactic acid fermentation is a common metabolic phenotype seen in most cancers including malignant gliomas. This metabolic phenotype is linked to abnormalities in mitochondrial structure and function. A luciferin-luciferase bioluminescence ATP assay was used to measure the influence of amino acids, glucose, and oxygen on ATP content and viability in mouse (VM-M3 and CT-2A) and human (U-87MG) glioma cells that differed in cell biology, genetic background, and species origin. Oxygen consumption was measured using the Resipher system. Extracellular lactate and succinate were measured as end products of the glycolysis and glutaminolysis pathways, respectively. The results showed that: (1) glutamine was a source of ATP content irrespective of oxygen. No other amino acid could replace glutamine in sustaining ATP content and viability; (2) ATP content persisted in the absence of glucose and under hypoxia, ruling out substantial contribution through either glycolysis or oxidative phosphorylation (OxPhos) under these conditions; (3) Mitochondrial complex IV inhibition showed that oxygen consumption was not an accurate measure for ATP production through OxPhos. The glutaminase inhibitor, 6-diazo-5-oxo-L-norleucine (DON), reduced ATP content and succinate export in cells grown in glutamine. The data suggests that mitochondrial substrate level phosphorylation in the glutamine-driven glutaminolysis pathway contributes to ATP content in these glioma cells. A new model is presented highlighting the synergistic interaction between the high-throughput glycolysis and glutaminolysis pathways that drive malignant glioma growth and maintain ATP content through the aerobic fermentation of both glucose and glutamine. LA - English DB - MTMT ER - TY - JOUR AU - Ravasz, Dóra AU - Bui, Dávid AU - Nazarian, Sara AU - Pallag, Gergely AU - Karnok, Noémi AU - Roberts, Jennie AU - Marzullo, Bryan P. AU - Tennant, Daniel A. AU - Greenwood, Bennett AU - Kitayev, Alex AU - Hill, Collin AU - Komlódi, Tímea AU - Doerrier, Carolina AU - Cunatova, Kristyna AU - Fernandez-Vizarra, Erika AU - Gnaiger, Erich AU - Kiebish, Michael A. AU - Raska, Alexandra AU - Kolev, Kraszimir Nikolaev AU - Czumbel, Bence AU - Narain, Niven R. AU - Seyfried, Thomas N. AU - Chinopoulos, Christos TI - Residual Complex I activity and amphidirectional Complex II operation support glutamate catabolism through mtSLP in anoxia JF - BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR BASIS OF DISEASE J2 - BBA-MOL BASIS DIS VL - 1870 PY - 2024 IS - 8 SP - 167388 SN - 0925-4439 DO - 10.1016/j.bbadis.2024.167388 UR - https://m2.mtmt.hu/api/publication/35626090 ID - 35626090 LA - English DB - MTMT ER - TY - JOUR AU - Raska, Alexandra AU - Balog Virág, Kata AU - Csikós, Petra (Metta) AU - Szabó, László AU - Kolev, Kraszimir Nikolaev AU - Wohner, Nikolett TI - Neutrophil Extracellular Traps: shaping fibrinolysis and fibrin structure in vivo and ex vivo JF - RESEARCH AND PRACTICE IN THROMBOSIS AND HAEMOSTASIS J2 - Res Pract Thromb Haemost VL - 8 PY - 2024 IS - Suppl. 2 SP - 76 EP - 77 PG - 2 SN - 2475-0379 UR - https://m2.mtmt.hu/api/publication/35595842 ID - 35595842 LA - English DB - MTMT ER - TY - JOUR AU - Balog Virág, Kata AU - B., Egri AU - Szabó, László AU - Kolev, Kraszimir Nikolaev AU - Wohner, Nikolett TI - The role of factor XIII in modulating the efficacy of tranexamic acid on fibrinolysis and fibrin structure in cellular and cell-free environments JF - RESEARCH AND PRACTICE IN THROMBOSIS AND HAEMOSTASIS J2 - Res Pract Thromb Haemost VL - 8 PY - 2024 IS - Suppl. 2 SP - 73 SN - 2475-0379 UR - https://m2.mtmt.hu/api/publication/35595783 ID - 35595783 LA - English DB - MTMT ER - TY - JOUR AU - Tóth, Balázs AU - Jiang, Yuefeng AU - Szöllősi, András AU - Zhang, Zhe AU - Csanády, László TI - A conserved mechanism couples cytosolic domain movements to pore gating in the TRPM2 channel JF - PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA J2 - P NATL ACAD SCI USA VL - 121 PY - 2024 IS - 46 PG - 12 SN - 0027-8424 DO - 10.1073/pnas.2415548121 UR - https://m2.mtmt.hu/api/publication/35594849 ID - 35594849 AB - Transient Receptor Potential Melastatin 2 (TRPM2) cation channels contribute to immunocyte activation, insulin secretion, and central thermoregulation. TRPM2 opens upon binding cytosolic Ca 2+ and ADP ribose (ADPR). We present here the 2.5 Å cryo-electronmicroscopy structure of TRPM2 from Nematostella vectensis (nvTRPM2) in a lipid nanodisc, complexed with Ca 2+ and ADPR-2′-phosphate. Comparison with nvTRPM2 without nucleotide reveals that nucleotide binding-induced movements in the protein’s three “core” layers deconvolve into a set of rigid-body rotations conserved from cnidarians to man. By covalently crosslinking engineered cysteine pairs we systematically trap the cytosolic layers in specific conformations and study effects on gate opening/closure. The data show that nucleotide binding in Layer 3 disrupts inhibitory intersubunit interactions, allowing rotation of Layer 2 which in turn expands the gate located in Layer 1. Channels trapped in that “activated” state are no longer nucleotide dependent, but are opened by binding of Ca 2+ alone. LA - English DB - MTMT ER - TY - JOUR AU - Raska, Alexandra AU - Baráth, Barbara AU - Balog Virág, Kata AU - Csikós, P AU - Szabó, László AU - Kolev, Kraszimir Nikolaev AU - Wohner, Nikolett TI - Neutrophil extracelluláris csapdák hatása a fibrinolízisre és fibrin szerkezetre in vivo és ex vivo JF - HEMATOLÓGIA-TRANSZFUZIOLÓGIA J2 - HEMATOLÓGIA-TRANSZFUZIOLÓGIA VL - 57 PY - 2024 IS - 3 SP - 187 EP - 188 PG - 2 SN - 1786-5913 UR - https://m2.mtmt.hu/api/publication/35573142 ID - 35573142 LA - Hungarian DB - MTMT ER - TY - JOUR AU - Komorowicz, Erzsébet AU - Gurabi, A AU - Sándor, Z AU - Wacha, András Ferenc AU - Kolev, Kraszimir Nikolaev TI - A natív és citrullinált core-hisztonok kringle-dependens módon gátolják a plazmin fibrin(ogeno)lítikus aktivitását JF - HEMATOLÓGIA-TRANSZFUZIOLÓGIA J2 - HEMATOLÓGIA-TRANSZFUZIOLÓGIA VL - 57 PY - 2024 IS - 3 SP - 183 EP - 184 PG - 2 SN - 1786-5913 DO - 10.1556/2068.2024.30001 UR - https://m2.mtmt.hu/api/publication/35573125 ID - 35573125 LA - Hungarian DB - MTMT ER - TY - JOUR AU - Balog Virág, Kata AU - Egri, B AU - Szabó, László AU - Kolev, Kraszimir Nikolaev AU - Wohner, Nikolett TI - A XIII-as faktor szerepe a tranexámsav fibrinolízisre és fibrinszerkezetre kifejtett hatásának modulálásában sejtes és sejtmentes környezetben JF - HEMATOLÓGIA-TRANSZFUZIOLÓGIA J2 - HEMATOLÓGIA-TRANSZFUZIOLÓGIA VL - 57 PY - 2024 IS - 3 SP - 172 SN - 1786-5913 UR - https://m2.mtmt.hu/api/publication/35573094 ID - 35573094 LA - Hungarian DB - MTMT ER - TY - JOUR AU - Mihályi, Csaba AU - Iordanov, Iordan AU - Szöllősi, András AU - Csanády, László TI - Structural determinants of protein kinase A essential for CFTR channel activation. JF - PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA J2 - P NATL ACAD SCI USA VL - 121 PY - 2024 IS - 46 PG - 11 SN - 0027-8424 DO - 10.1073/pnas.2407728121 UR - https://m2.mtmt.hu/api/publication/35501499 ID - 35501499 AB - Cystic Fibrosis Transmembrane Conductance Regulator (CFTR), the anion channel mutated in cystic fibrosis (CF) patients, is activated by the catalytic subunit of protein kinase A (PKA-C). PKA-C activates CFTR both noncatalytically, through binding, and catalytically, through phosphorylation of multiple serines in CFTR's regulatory (R) domain. Here, we identify key molecular determinants of the CFTR/PKA-C interaction essential for these processes. By comparing CFTR current activation in the presence of ATP or an ATP analog unsuitable for phosphotransfer, as well as pseudosubstrate peptides of various lengths, we identify two distinct specific regions of the PKA-C surface which interact with CFTR to cause noncatalytic and catalytic CFTR stimulation, respectively. Whereas the "substrate site" mediates CFTR phosphorylation, a distinct hydrophobic patch (the "docking site") is responsible for noncatalytic CFTR activation, achieved by stabilizing the R domain in a "released" conformation permissive to channel gating. Furthermore, by comparing PKA-C variants with different posttranslational modification patterns, we find that direct membrane tethering of the kinase through its N-terminal myristoyl group is an unappreciated fundamental requirement for CFTR activation: PKA-C demyristoylation abolishes noncatalytic, and profoundly slows catalytic, CFTR stimulation. For the F508del CFTR mutant, present in ~90% of CF patients, maximal activation by demyristoylated PKA-C is reduced by ~10-fold compared to that by myristoylated PKA-C. Finally, in bacterial genera that contain common CF pathogens, we identify virulence factors that demyristoylate PKA-C in vitro, raising the possibility that during recurrent bacterial infections in CF patients, PKA-C demyristoylation may contribute to the exacerbation of lung disease. LA - English DB - MTMT ER - TY - JOUR AU - Fiedorczuk, Karol AU - Iordanov, Iordan AU - Mihályi, Csaba AU - Szöllősi, András AU - Csanády, László AU - Chen, Jue TI - The structures of protein kinase A in complex with CFTR. Mechanisms of phosphorylation and noncatalytic activation. TS - Mechanisms of phosphorylation and noncatalytic activation. JF - PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA J2 - P NATL ACAD SCI USA VL - 121 PY - 2024 IS - 46 PG - 10 SN - 0027-8424 DO - 10.1073/pnas.2409049121 UR - https://m2.mtmt.hu/api/publication/35501481 ID - 35501481 AB - Protein kinase A (PKA) is a key regulator of cellular functions by selectively phosphorylating numerous substrates, including ion channels, enzymes, and transcription factors. It has long served as a model system for understanding the eukaryotic kinases. Using cryoelectron microscopy, we present complex structures of the PKA catalytic subunit (PKA-C) bound to a full-length protein substrate, the cystic fibrosis transmembrane conductance regulator (CFTR)-an ion channel vital to human health. CFTR gating requires phosphorylation of its regulatory (R) domain. Unphosphorylated CFTR engages PKA-C at two locations, establishing two "catalytic stations" near to, but not directly involving, the R domain. This configuration, coupled with the conformational flexibility of the R domain, permits transient interactions of the eleven spatially separated phosphorylation sites. Furthermore, we determined two structures of the open-pore CFTR stabilized by PKA-C, providing a molecular basis to understand how PKA-C stimulates CFTR currents even in the absence of phosphorylation. LA - English DB - MTMT ER -