Characterization of surface structure and p47phox SH3 domain-mediated conformational changes for human neutrophil flavocytochrome b.
Characterization of surface structure and p47phox SH3 domain-mediated conformational changes for human neutrophil flavocytochrome b.
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人中性粒细胞黄素细胞色素 b 的表面结构和 p47phox SH3 结构域介导的构象变化的表征。
DOI：
10.1021/bi701626p
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发表时间：
2007
期刊：
影响因子：
2.9
通讯作者：
Jesaitis,AlgirdasJ
中科院分区：
文献类型：
--
作者：
Taylor,RossM;Lord,ConnieI;Riesselman,MarciaH;Gripentrog,JeannieM;Leto,ThomasL;McPhail,LindaC;Berdichevsky,Yevgeny;Pick,Edgar;Jesaitis,AlgirdasJ
异二聚体的膜蛋白黄细胞色素b（Cytb）是吞噬细胞NADPH氧化酶的催化核心，并产生在宿主防御中起关键作用的超氧化物。为了更好地定义这种多亚基酶复合物的超氧化物生成的激活，Cytb特异性单克隆抗体（mAb）和p47 phoxSH 3结构域（p47 SH 3AB）在本研究中被用作探针，以映射人中性粒细胞Cytb的表面结构和构象动力学。在与洗涤剂溶解的Cytb的下拉和共免疫沉淀研究中，氧化酶抑制性mAb CS 9显示与p47 SH 3AB在p22 phox亚基的C-末端区域上共享重叠结合位点。类似的研究表明，mAb 44.1和CS 9/p47 SH 3AB结合位点之间令人惊讶地缺乏重叠，并且它们表明氧化酶抑制性mAb NL 7结合与p22 phoxC末端结构域物理分离的区域。共振能量转移和尺寸排阻色谱法证实了功能性重构Cytb带的上述结果，提供了mAb CS 9和p47 SH 3AB两者的结合改变Cytb构象的证据。进一步支持p47 phoxSH 3结构域的结合调节Cytb的结构的实验是使用无细胞测定系统获得的，其中在p67 phox（1 - 212）-Rac 1（Q61 L）融合蛋白存在下，p47 SH 3AB增强了超氧化物的产生。总之，本研究进一步表征了人中性粒细胞Cytb的结构，包括洗涤剂胶束和重建的膜双层，并提供了证据表明，在氧化酶活化过程中，胞质调节亚基p47 phox调节Cytb的构象（除了作为衔接蛋白）。
The heterodimeric, integral membrane protein flavocytochromeb(Cytb) is the catalytic core of the phagocyte NADPH oxidase and generates superoxide which plays a critical role in host defense. To better define the activation of superoxide production by this multisubunit enzyme complex, Cytb-specific monoclonal antibodies (mAbs) and the p47phoxSH3 domains (p47SH3AB) were used in the present study as probes to map surface structure and conformational dynamics in human neutrophil Cytb. In pull-down and co-immunoprecipitation studies with detergent-solubilized Cytb, the oxidase-inhibitory mAb CS9 was shown to share an overlapping binding site with p47SH3ABon the C-terminal region of the p22phoxsubunit. Similar studies demonstrated a surprising lack of overlap between the mAb 44.1 and CS9/p47SH3ABbinding sites, and they indicated that the oxidase-inhibitory mAb NL7 binds a region physically separated from the p22phoxC-terminal domain. Resonance energy transfer and size exclusion chromatography confirmed the above results for functionally reconstituted Cytband provided evidence that binding of both mAb CS9 and p47SH3ABaltered the conformation of Cytb. Further support that binding of the p47phoxSH3 domains modulates the structure of Cytbwas obtained using a cell-free assay system where p47SH3ABenhanced superoxide production in the presence of a p67phox(1−212)-Rac1(Q61L) fusion protein. Taken together, this study further characterizes the structure of human neutrophil Cytbin both detergent micelles and reconstituted membrane bilayers, and it provides evidence that the cytosolic regulatory subunit p47phoxmodulates the conformation of Cytb(in addition to serving as an adapter protein) during oxidase activation.
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影响因子：
15
作者：
Krieger, F;Möglich, A;Kiefhaber, T
通讯作者：
Kiefhaber, T
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
NADPH 氧化酶活性的位点特异性抑制剂和黄细胞色素 b 的结构探针：针对 p22phox 亚基 1 的六种单克隆抗体的表征
影响因子：
4.4
作者：
Ross M. Taylor;J. B. Burritt;D. Baniulis;T. R. Foubert;Connie I. Lord;M. Dinauer;C. Parkos;A. J. Jesaitis
通讯作者：
A. J. Jesaitis
影响因子：
56.9
作者：
ROTROSEN, D;YEUNG, CL;KWONG, CH
通讯作者：
KWONG, CH
影响因子：
3.9
作者：
Campion, Yannick;Paclet, Marie-Helene;Morel, Francoise
通讯作者：
Morel, Francoise