Nox5 forms a functional oligomer mediated by self-association of its dehydrogenase domain.
Nox5 forms a functional oligomer mediated by self-association of its dehydrogenase domain.
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NOX5形成了通过其脱氢酶结构域的自我关联介导的功能性低聚物。
DOI：
10.1021/bi1020088
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发表时间：
2011-03-29
期刊：
影响因子：
2.9
通讯作者：
Lambeth JD
中科院分区：
文献类型：
--
作者：
Kawahara T;Jackson HM;Smith SM;Simpson PD;Lambeth JD
Nox5 belongs to the calcium-regulated subfamily of NADPH oxidases (Nox). Like other calcium-regulated Noxes, Nox5 has an EF hand-containing calcium-binding domain at its N-terminus, a transmembrane heme-containing region and a C-terminal dehydrogenase (DH) domain that binds FAD and NADPH. While Nox1-4 require regulatory subunits including p22phox, Nox5 activity does not depend on any subunits. We found that inactive point mutants and truncated forms of Nox5 (including the naturally expressed splice form Nox5S) inhibit full-length Nox5, consistent with formation of a dominant negative complex. Oligomerization of full-length Nox5 was demonstrated using co-immunoprecipitation of co-expressed, differentially tagged forms of Nox5 and occurred independently of calcium ion. Several approaches were used to show that the DH domain mediates oligomerization: Nox5 could be isolated as a multimer when the calcium-binding domain and/or the N-terminal polybasic region (PBR-N) were deleted, but deletion of the DH domain eliminated oligomerization. Further, a chimera containing the transmembrane domain of Ciona intestinalis voltage sensor-containing phosphatase (CiVSP) fused to the Nox5 DH domain formed a co-immunoprecipitating complex with, and functioned as a dominant inhibitor of, full-length Nox5. Radiation inactivation of Nox5 overexpressed in HEK293 cells or endogenously expressed in human aortic smooth muscle cells indicated molecular weights of about 350 kDa and 300 kDa, respectively, consistent with a tetramer as the functionally active unit. Thus, Nox5 forms a catalytically active oligomer in the membrane that is mediated by its dehydrogenase domain. As a result of oligomerization, the short, calcium-independent splice form Nox5S may function as an endogenous inhibitor of calcium-stimulated ROS generation by full-length Nox5.
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影响因子：
2.9
作者：
BEAUREGARD, G;POTIER, M
通讯作者：
POTIER, M
影响因子：
7.4
作者：
Jay, Desmond B.;Papaharalambus, Christopher A.;Griendling, Kathy K.
通讯作者：
Griendling, Kathy K.
影响因子：
4.8
作者：
Grasberger, Helmut;Refetoff, Samuel
通讯作者：
Refetoff, Samuel
影响因子：
3.3
作者：
Kawahara, Tsukasa;Lambeth, J. David
通讯作者：
Lambeth, J. David
DOI：
10.1016/0005-2736(68)90097-7
发表时间：
1968-01-01
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA
影响因子：
--
作者：
KEPNER, GR;MACEY, RI
通讯作者：
MACEY, RI