TY - JOUR AU - Iordanov, Iordan AU - Tóth, Balázs AU - Szöllősi, András AU - Csanády, László TI - Enzyme activity and selectivity filter stability of ancient TRPM2 channels were simultaneously lost in early vertebrates JF - ELIFE J2 - ELIFE VL - 8 PY - 2019 IS - 2019 PG - 23 SN - 2050-084X DO - 10.7554/eLife.44556 UR - https://m2.mtmt.hu/api/publication/30637149 ID - 30637149 N1 - Funding Agency and Grant Number: Howard Hughes Medical InstituteHoward Hughes Medical Institute; Magyar Tudomanyos Akademia [LP2017-14/2017]; Ministry of Human Capacities of Hungary [UNKP 17-4-I-SE-61, UNKP 18-4-SE-132]; Magyar Tudomanyos Akademia Funding text: Howard Hughes Medical Institute International Early Career Scientist Award Laszlo Csanady; Magyar Tudomanyos Akademia LP2017-14/2017 Laszlo Csanady; Ministry of Human Capacities of Hungary UNKP 17-4-I-SE-61 Balazs Toth; Magyar Tudomanyos Akademia Bolyai Research Fellowship Balazs Toth; Ministry of Human Capacities of Hungary UNKP-FIKP Laszlo Csanady; Ministry of Human Capacities of Hungary UNKP 18-4-SE-132 Balazs Toth; The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. Export Date: 7 January 2020 Correspondence Address: Csanády, L.; Department of Medical Biochemistry, Semmelweis UniversityHungary; email: csanady.laszlo@med.semmelweis-univ.hu Export Date: 8 January 2020 Correspondence Address: Csanády, L.; Department of Medical Biochemistry, Semmelweis UniversityHungary; email: csanady.laszlo@med.semmelweis-univ.hu AB - Transient Receptor Potential Melastatin 2 (TRPM2) is a cation channel important for the immune response, insulin secretion, and body temperature regulation. It is activated by cytosolic ADP ribose (ADPR) and contains a nudix-type motif 9 (NUDT9)-homology (NUDT9-H) domain homologous to ADPR phosphohydrolases (ADPRases). Human TRPM2 (hsTRPM2) is catalytically inactive due to mutations in the conserved Nudix box sequence. Here, we show that TRPM2 Nudix motifs are canonical in all invertebrates but vestigial in vertebrates. Correspondingly, TRPM2 of the cnidarian Nematostella vectensis (nvTRPM2) and the choanoflagellate Salpingoeca rosetta (srTRPM2) are active ADPRases. Disruption of ADPRase activity fails to affect nvTRPM2 channel currents, reporting a catalytic cycle uncoupled from gating. Furthermore, pore sequence substitutions responsible for inactivation of hsTRPM2 also appeared in vertebrates. Correspondingly, zebrafish (Danio rerio) TRPM2 (drTRPM2) and hsTRPM2 channels inactivate, but srTRPM2 and nvTRPM2 currents are stable. Thus, catalysis and pore stability were lost simultaneously in vertebrate TRPM2 channels. LA - English DB - MTMT ER - TY - JOUR AU - Liu, F AU - Zhang, Z AU - Csanády, László AU - Gadsby, DC AU - Chen, J TI - Molecular Structure of the Human CFTR Ion Channel JF - CELL J2 - CELL VL - 169 PY - 2017 IS - 1 SP - 85 EP - 95.e8 SN - 0092-8674 DO - 10.1016/j.cell.2017.02.024 UR - https://m2.mtmt.hu/api/publication/3210856 ID - 3210856 N1 - Cited By :244 Export Date: 9 March 2022 CODEN: CELLB Correspondence Address: Chen, J.; Laboratory of Membrane Biophysics and Biology, 1230 York Avenue, United States Chemicals/CAS: adenosine triphosphatase, 37289-25-1, 9000-83-3; adenosine triphosphate, 15237-44-2, 56-65-5, 987-65-5; arginine, 1119-34-2, 15595-35-4, 7004-12-8, 74-79-3; cyclic AMP dependent protein kinase; cysteine, 4371-52-2, 52-89-1, 52-90-4; cystic fibrosis transmembrane conductance regulator, 126880-72-6; lysine, 56-87-1, 6899-06-5, 70-54-2; multidrug resistance associated protein 1; proline, 147-85-3, 7005-20-1; protein, 67254-75-5; serine, 56-45-1, 6898-95-9; Adenosine Triphosphate; CFTR protein, human; CFTR protein, zebrafish; Cystic Fibrosis Transmembrane Conductance Regulator; Zebrafish Proteins Funding details: LP2012-39/2012 Funding details: Howard Hughes Medical Institute, HHMI Funding details: Cystic Fibrosis Foundation, CFF, CSANAD15G0 Funding details: Rockefeller University Funding text 1: We thank Eric Gouaux for the expression vector, Mark Ebrahim and Johanna Sotiris at the Rockefeller Evelyn Gruss Lipper Cryo-Electron Microscopy Resource Center for assistance in data collection, and Sarah McCarry for editing this manuscript. This work is supported by the Rockefeller University (to J.C. and D.C.G), the Howard Hughes Medical Institute (to J.C.), and MTA-Momentum (LP2012-39/2012) and Cystic Fibrosis Foundation (CSANAD15G0) grants (to L.C.). AB - The cystic fibrosis transmembrane conductance regulator (CFTR) is an ATP-binding cassette (ABC) transporter that uniquely functions as an ion channel. Here, we present a 3.9 Å structure of dephosphorylated human CFTR without nucleotides, determined by electron cryomicroscopy (cryo-EM). Close resemblance of this human CFTR structure to zebrafish CFTR under identical conditions reinforces its relevance for understanding CFTR function. The human CFTR structure reveals a previously unresolved helix belonging to the R domain docked inside the intracellular vestibule, precluding channel opening. By analyzing the sigmoid time course of CFTR current activation, we propose that PKA phosphorylation of the R domain is enabled by its infrequent spontaneous disengagement, which also explains residual ATPase and gating activity of dephosphorylated CFTR. From comparison with MRP1, a feature distinguishing CFTR from all other ABC transporters is the helix-loop transition in transmembrane helix 8, which likely forms the structural basis for CFTR's channel function. © 2017 Elsevier Inc. 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 - Csanády, László AU - Seto-Young, D AU - Chan, KW AU - Cenciarelli, C AU - Angel, BB AU - Qin, J AU - McLachlin, DT AU - Krutchinsky, AN AU - Chait, BT AU - Nairn, AC AU - Gadsby, DC TI - Preferential phosphorylation of R-domain serine 768 dampens activation of CFTR channels by PKA JF - JOURNAL OF GENERAL PHYSIOLOGY J2 - J GEN PHYSIOL VL - 125 PY - 2005 IS - 2 SP - 171 EP - 186 PG - 16 SN - 0022-1295 DO - 10.1085/jgp.200409076 UR - https://m2.mtmt.hu/api/publication/1493089 ID - 1493089 LA - English DB - MTMT ER - TY - JOUR AU - Chan, KW AU - Csanády, László AU - Seto-Young, D AU - Nairn, AC AU - Gadsby, DC TI - Severed molecules functionally define the boundaries of the cystic fibrosis transmembrane conductance regulator's NH2-terminal nucleotide binding domain JF - JOURNAL OF GENERAL PHYSIOLOGY J2 - J GEN PHYSIOL VL - 116 PY - 2000 IS - 2 SP - 163 EP - 180 PG - 18 SN - 0022-1295 DO - 10.1085/jgp.116.2.163 UR - https://m2.mtmt.hu/api/publication/1493091 ID - 1493091 N1 - Összes idézések száma a WoS-ban: 0 LA - English DB - MTMT ER - TY - JOUR AU - Csanády, László AU - Chan, KW AU - Seto-Young, D AU - Kopsco, DC AU - Nairn, AC AU - Gadsby, DC TI - Severed channels probe regulation of gating of cystic fibrosis transmembrane conductance regulator by its cytoplasmic domains JF - JOURNAL OF GENERAL PHYSIOLOGY J2 - J GEN PHYSIOL VL - 116 PY - 2000 IS - 3 SP - 477 EP - 500 PG - 24 SN - 0022-1295 DO - 10.1085/jgp.116.3.477 UR - https://m2.mtmt.hu/api/publication/1493767 ID - 1493767 N1 - Laboratory of Cardiac/Membrane Physiology, Rockefeller University, New York, NY 10021-6399, United States Laboratory of Molecular and Cellular Neuroscience, Rockefeller University, New York, NY 10021-6399, United States Laboratory of Cardiac/Membrane Physiology, Rockefeller University, 1230 York Avenue, New York, NY 10021-6399, United States Cited By :99 Export Date: 27 February 2023 CODEN: JGPLA Correspondence Address: Gadsby, D.C.; Lab. of Cardiac/Membrane Physiology, 1230 York Avenue, New York, NY 10021-6399, United States; email: gadsby@rockvax.rockefeller.edu Chemicals/CAS: Adenosine Triphosphate, 56-65-5; Adenylyl Imidodiphosphate, 25612-73-1; CFTR protein, human; Cyclic AMP-Dependent Protein Kinases, EC 2.7.1.37; Cystic Fibrosis Transmembrane Conductance Regulator, 126880-72-6; DNA Primers; Recombinant Proteins Funding details: National Institute of Diabetes and Digestive and Kidney Diseases, NIDDK, R01DK051767 LA - English DB - MTMT ER -