Regulation of the human neutrophil NADPH oxidase by rho-related G-proteins.
Regulation of the human neutrophil NADPH oxidase by rho-related G-proteins.
复制标题
rho 相关 G 蛋白对人中性粒细胞 NADPH 氧化酶的调节。
作者：
C. H. Kwong;Harry L Malech;D. Rotrosen;T. Leto
吞噬性白色血细胞产生超氧化物需要从膜和胞质蛋白组装NADPH氧化酶。重组胞质蛋白p47 phox和p67 phox和中性粒细胞膜用于纯化NADPH氧化酶无细胞重建所必需和足够的第三种胞质组分。该组分被分离为rho-GDP解离抑制剂（rho-GDI）和ras相关鸟嘌呤核苷酸结合蛋白rho亚家族的两个成员rac 2和CDC 42 Hs的复合物。氧化酶重建与这些纯的胞质蛋白不受GTP γ S的影响，但被GDP β S抑制，表明活性复合物含有内源性结合GTP。在与放射性标记的鸟嘌呤核苷酸交换后，通过凝胶过滤证明rho-GDI与这些G蛋白的GTP γ S结合形式的直接结合。在比较这些G蛋白的纯重组形式的活性的实验中，rho-GDI被证明对于无细胞氧化酶重建是不必要的。重组外消旋体增加超氧化物的产生，而与外消旋体共享70%氨基酸序列同一性的重组CDC 42 H则没有。rac 1和rac 2的三个高度保守的区域在CDC 42 H中被注意到为明显不同。有人提出，一个或多个这些区域的外消旋可能涉及的外消旋与其他NADPH氧化酶蛋白的特异性相互作用。
Superoxide production by phagocytic white blood cells requires the assembly of an NADPH oxidase from membrane and cytosolic proteins. Recombinant cytosolic proteins p47phox and p67phox and neutrophil membranes were used to purify a third cytosolic component that is necessary and sufficient for cell-free reconstitution of NADPH oxidase. The component was isolated as a complex of rho-GDP dissociation inhibitor (rho-GDI) and two members of the rho subfamily of ras-related guanine nucleotide binding proteins, rac2 and CDC42Hs. Oxidase reconstitution with these pure cytosolic proteins was unaffected by GTP gamma S but was inhibited by GDP beta S, suggesting that the active complex contained endogenous bound GTP. Direct binding of rho-GDI to the GTP gamma S-bound forms of these G-proteins was demonstrated by gel filtration following exchange with radiolabeled guanine nucleotide. rho-GDI was shown to be nonessential for cell-free oxidase reconstitution in experiments that compared the activities of pure recombinant forms of these G-proteins. Recombinant rac augmented superoxide production, while recombinant CDC42Hs, which shares 70% amino acid sequence identity with rac, did not. Three highly conserved regions of rac1 and rac2 were noted as markedly divergent in CDC42Hs. It is proposed that one or more of these regions of rac may be involved in the specific interaction of rac with the other NADPH oxidase protein(s).
登录
查看更多内容
影响因子：
56.9
作者：
LETO, TL;LOMAX, KJ;MALECH, HL
通讯作者：
MALECH, HL
DOI：
10.1073/pnas.84.20.6970
发表时间：
1987-10-01
影响因子：
11.1
作者：
AEBERSOLD, RH;LEAVITT, J;KENT, SBH
通讯作者：
KENT, SBH
常染色体慢性肉芽肿病中不存在两种胞质中性粒细胞氧化酶成分。
DOI：
10.1126/science.2848318
发表时间：
1988
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Volpp,BD;Nauseef,WM;Clark,RA
通讯作者：
Clark,RA
DOI：
10.1073/pnas.87.24.9853
发表时间：
1990-12
影响因子：
11.1
作者：
K. Shinjo;J. Koland;Matthew J. Hart;Vikram Narasimhan;Douglas I. Johnson;T. Evans;R. Cerione
通讯作者：
K. Shinjo;J. Koland;Matthew J. Hart;Vikram Narasimhan;Douglas I. Johnson;T. Evans;R. Cerione
影响因子：
4.1
作者：
Gibson, RM;Wilson-Delfosse, AL
通讯作者：
Wilson-Delfosse, AL