dc.contributor.author |
Iordanov, Iordan |
|
dc.contributor.author |
Mihályi, Csaba |
|
dc.contributor.author |
Tóth, Balázs |
|
dc.contributor.author |
Csanády, László |
|
dc.date.accessioned |
2018-10-01T09:42:02Z |
|
dc.date.available |
2018-10-01T09:42:02Z |
|
dc.date.issued |
2016 |
|
dc.identifier.citation |
pagination=e17600, 20 pages;journalVolume=5;journalTitle=ELIFE; |
hu |
dc.identifier.uri |
http://repo.lib.semmelweis.hu//handle/123456789/6050 |
|
dc.identifier.uri |
doi:10.7554/eLife.17600 |
|
dc.description.abstract |
Transient Receptor Potential Melastatin 2 (TRPM2) is a Ca(2+)-permeable cation channel essential for immunocyte activation, insulin secretion, and postischemic cell death. TRPM2 is activated by ADP ribose (ADPR) binding to its C-terminal cytosolic NUDT9-homology (NUDT9H) domain, homologous to the soluble mitochondrial ADPR pyrophosphatase (ADPRase) NUDT9. Reported ADPR hydrolysis classified TRPM2 as a channel-enzyme, but insolubility of isolated NUDT9H hampered further investigations. Here we developed a soluble NUDT9H model using chimeric proteins built from complementary polypeptide fragments of NUDT9H and NUDT9. When expressed in E.coli, chimeras containing up to ~90% NUDT9H sequence remained soluble and were affinity-purified. In ADPRase assays the conserved Nudix-box sequence of NUDT9 proved essential for activity (kcat~4-9s(-1)), that of NUDT9H did not support catalysis. Replacing NUDT9H in full-length TRPM2 with soluble chimeras retained ADPR-dependent channel gating (K1/2~1-5 μM), confirming functionality of chimeric domains. Thus, TRPM2 is not a 'chanzyme'. Chimeras provide convenient soluble NUDT9H models for structural/biochemical studies. |
|
dc.relation.ispartof |
urn:issn:2050-084X |
|
dc.title |
The proposed channel-enzyme transient receptor potential melastatin 2 does not possess ADP ribose hydrolase activity |
hu |
dc.type |
Journal Article |
hu |
dc.date.updated |
2018-08-08T10:03:41Z |
|
dc.language.rfc3066 |
en |
hu |
dc.identifier.mtmt |
3105008 |
|
dc.identifier.wos |
000380856900001 |
|
dc.identifier.pubmed |
27383051 |
|
dc.contributor.department |
SE/AOK/I/Orvosi Biokémiai Intézet |
|
dc.contributor.institution |
Semmelweis Egyetem |
|
dc.mtmt.swordnote |
FELTÖLTŐ: Bökönyi Zita - bokonyi.zita@med.semmelweis-univ.hu |
|