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 - Zhang, Zhe AU - Tóth, Balázs AU - Szöllősi, András AU - Chen, Jue AU - Csanády, László TI - Structure of a TRPM2 channel in complex with Ca2+ explains unique gating regulation JF - ELIFE J2 - ELIFE VL - 7 PY - 2018 PG - 22 SN - 2050-084X DO - 10.7554/eLife.36409 UR - https://m2.mtmt.hu/api/publication/30465914 ID - 30465914 AB - Transient receptor potential melastatin 2 (TRPM2) is a Ca2+-permeable cation channel required for immune cell activation, insulin secretion, and body heat control. TRPM2 is activated by cytosolic Ca2+, phosphatidyl-inositol-4,5-bisphosphate and ADP ribose. Here, we present the 3 A resolution electron cryo-microscopic structure of TRPM2 from Nematostella vectensis, 63% similar in sequence to human TRPM2, in the Ca2+-bound closed state. Compared to other TRPM channels, TRPM2 exhibits unique structural features that correlate with its function. The pore is larger and more negatively charged, consistent with its high Ca2+ selectivity and larger conductance. The intracellular Ca2+ binding sites are connected to the pore and cytosol, explaining the unusual dependence of TRPM2 activity on intra- and extracellular Ca2+. In addition, the absence of a post filter motif is likely the cause of the rapid inactivation of human TRPM2. Together, our cryo-EM and electrophysiology studies provide a molecular understanding of the unique gating mechanism of TRPM2. LA - English DB - MTMT ER - TY - JOUR AU - Iordanov, Iordan AU - Mihályi, Csaba AU - Tóth, Balázs AU - Csanády, László TI - The proposed channel-enzyme transient receptor potential melastatin 2 does not possess ADP ribose hydrolase activity JF - ELIFE J2 - ELIFE VL - 5 PY - 2016 PG - 20 SN - 2050-084X DO - 10.7554/eLife.17600 UR - https://m2.mtmt.hu/api/publication/3105008 ID - 3105008 N1 - Department of Medical Biochemistry, Semmelweis University, Budapest, Hungary MTA-SE Ion Channel Research Group, Semmelweis University, Budapest, Hungary Cited By :17 Export Date: 20 September 2019 Correspondence Address: Csanády, L.; Department of Medical Biochemistry, Semmelweis UniversityHungary; email: csanady.laszlo@med.semmelweis-univ.hu Funding Agency and Grant Number: Howard Hughes Medical Institute International Early Career Scientist [55007416]; Magyar Tudomanyos Akademia Lendulet [LP2012-39/2012] Funding text: Howard Hughes Medical Institute International Early Career Scientist grant, 55007416 Laszlo Csanady; Magyar Tudomanyos Akademia Lendulet grant, LP2012-39/2012 Laszlo Csanady Export Date: 7 January 2020 Correspondence Address: Csanády, L.; Department of Medical Biochemistry, Semmelweis UniversityHungary; email: csanady.laszlo@med.semmelweis-univ.hu LA - English DB - MTMT ER - TY - JOUR AU - Tóth, Balázs AU - Iordanov, Iordan AU - Csanády, László TI - Ruling out pyridine dinucleotides as true TRPM2 channel activators reveals novel direct agonist ADP-ribose-2'-phosphate JF - JOURNAL OF GENERAL PHYSIOLOGY J2 - J GEN PHYSIOL VL - 145 PY - 2015 IS - 5 SP - 419 EP - 430 PG - 12 SN - 0022-1295 DO - 10.1085/jgp.201511377 UR - https://m2.mtmt.hu/api/publication/2949645 ID - 2949645 N1 - Funding Agency and Grant Number: International Early Career Scientist grant from the Howard Hughes Medical InstituteHoward Hughes Medical Institute; MTA Lendulet grant [LP2012-39/2012] Funding text: This work is supported by an International Early Career Scientist grant from the Howard Hughes Medical Institute to L. Csanady, and MTA Lendulet grant LP2012-39/2012. Export Date: 7 January 2020 CODEN: JGPLA Correspondence Address: Csanády, L.; Department of Medical Biochemistry, Semmelweis UniversityHungary; email: csanay.laszlo@med.semmelweis-univ.hu AB - Transient receptor potential melastatin 2 (TRPM2), a Ca(2+)-permeable cation channel implicated in postischemic neuronal cell death, leukocyte activation, and insulin secretion, is activated by intracellular ADP ribose (ADPR). In addition, the pyridine dinucleotides nicotinamide-adenine-dinucleotide (NAD), nicotinic acid-adenine-dinucleotide (NAAD), and NAAD-2'-phosphate (NAADP) have been shown to activate TRPM2, or to enhance its activation by ADPR, when dialyzed into cells. The precise subset of nucleotides that act directly on the TRPM2 protein, however, is unknown. Here, we use a heterologously expressed, affinity-purified-specific ADPR hydrolase to purify commercial preparations of pyridine dinucleotides from substantial contaminations by ADPR or ADPR-2'-phosphate (ADPRP). Direct application of purified NAD, NAAD, or NAADP to the cytosolic face of TRPM2 channels in inside-out patches demonstrated that none of them stimulates gating, or affects channel activation by ADPR, indicating that none of these dinucleotides directly binds to TRPM2. Instead, our experiments identify for the first time ADPRP as a true direct TRPM2 agonist of potential biological interest. LA - English DB - MTMT ER - TY - JOUR AU - Tóth, Balázs AU - Csanády, László TI - Pore collapse underlies irreversible inactivation of TRPM2 cation channel currents. JF - PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA J2 - P NATL ACAD SCI USA VL - 109 PY - 2012 IS - 33 SP - 13440 EP - 13445 PG - 6 SN - 0027-8424 DO - 10.1073/pnas.1204702109 UR - https://m2.mtmt.hu/api/publication/2041974 ID - 2041974 N1 - Funding Agency and Grant Number: Orszagos Tudomanyos Kutatasi Alapprogramok Grant [F 68143]; Howard Hughes Medical InstituteHoward Hughes Medical Institute Funding text: We thank Dorottya Mayer for oocyte isolation and injection; Tibor Rohacs for the TRPM8 clone, for PIP2, and for much valuable advice; and David Gadsby for discussions. L. C. is a Bolyai Research Fellow of the Hungarian Academy of Sciences. Support for this work was provided by Orszagos Tudomanyos Kutatasi Alapprogramok Grant F 68143 (to L. C.) and an International Early Career Scientist grant from the Howard Hughes Medical Institute (to L.C.). Cited By :33 Export Date: 7 January 2020 CODEN: PNASA Correspondence Address: Csanády, L.; Department of Medical Biochemistry, Semmelweis University, Budapest H-1094, Hungary; email: csanady.laszlo@med.semmelweis-univ.hu AB - The Ca(2+)-permeable cation channel transient receptor potential melastatin 2 (TRPM2) plays a key role in pathogen-evoked phagocyte activation, postischemic neuronal apoptosis, and glucose-evoked insulin secretion, by linking these cellular responses to oxidative stress. TRPM2 channels are coactivated by binding of intracellular ADP ribose and Ca(2+) to distinct cytosolically accessible sites on the channels. These ligands likely regulate the activation gate, conserved in the voltage-gated cation channel superfamily, that comprises a helix bundle formed by the intracellular ends of transmembrane helix six of each subunit. For several K(+) and TRPM family channels, activation gate opening requires the presence of phosphatidylinositol-bisphosphate (PIP(2)) in the inner membrane leaflet. Most TRPM family channels inactivate upon prolonged stimulation in inside-out patches; this "rundown" is due to PIP(2) depletion. TRPM2 currents also run down within minutes, but the molecular mechanism of this process is unknown. Here we report that high-affinity PIP(2) binding regulates Ca(2+) sensitivity of TRPM2 activation. Nevertheless, TRPM2 inactivation is not due to PIP(2) depletion; rather, it is state dependent, sensitive to permeating ions, and can be completely prevented by mutations in the extracellular selectivity filter. Introduction of two negative charges plus a single-residue insertion, to mimic the filter sequence of TRPM5, results in TRPM2 channels that maintain unabated maximal activity for over 1 h, and display altered permeation properties but intact ADP ribose/Ca(2+)-dependent gating. Thus, upon prolonged stimulation, the TRPM2 selectivity filter undergoes a conformational change reminiscent of that accompanying C-type inactivation of voltage-gated K(+) channels. The noninactivating TRPM2 variant will be invaluable for gating studies. 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 -