Molecular cloning and expression of a cDNA encoding endothelial cell nitric oxide synthase.
Molecular cloning and expression of a cDNA encoding endothelial cell nitric oxide synthase.
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DOI：
10.1016/s0021-9258(19)49528-8
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发表时间：
1992-08
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影响因子：
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通讯作者：
W. Sessa;Jeffrey K. Harrison;C. Barber;D. Zeng;M. Durieux;D. D. D’Angelo-D.;K. Lynch;M. Peach
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文献类型：
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作者：
W. Sessa;Jeffrey K. Harrison;C. Barber;D. Zeng;M. Durieux;D. D. D’Angelo-D.;K. Lynch;M. Peach
Endothelium-derived relaxing factor (EDRF), identified as nitric oxide (NO), is derived from a guanidino nitrogen of L-arginine via its metabolism by nitric oxide synthase (NOS). Herein, we report the molecular cloning of a cDNA encoding the constitutive calcium-calmodulin (Ca2+/CaM)-regulated nitric oxide synthase (ECNOS). A full-length ECNOS clone was isolated by screening a bovine aortic endothelial cell cDNA library using a fragment of rat brain NOS (bNOS) cDNA. This cDNA has an open reading frame of 3615 nucleotides encoding a 1205-amino acid protein. Membranes prepared from COS cells transfected with the ECNOS cDNA demonstrated NADPH- and Ca2+/CaM- dependent conversion of L-, but not D-, arginine to NO and citrulline that was inhibited by NG-nitro-L-arginine methyl ester. Comparison of the deduced amino acid sequence of ECNOS to the bNOS and macrophage NOS (Mac-NOS) sequences revealed 57 and 50% identity, respectively. In addition, ECNOS contains a unique N-myristylation consensus sequence (not shared by bNOS or Mac-NOS) that may explain its membrane localization.
