Identification of cytochrome P450 2C2 protein complexes in mouse liver.
Identification of cytochrome P450 2C2 protein complexes in mouse liver.
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DOI：
10.1002/pmic.201100001
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发表时间：
2011-08
期刊：
影响因子：
3.4
通讯作者：
Kemper, Byron
中科院分区：
文献类型：
--
作者：
Li, Bin;Yau, Peter;Kemper, Byron
关键词：
微粒体细胞色素P450（CYP）与微粒体膜上其他蛋白质的相互作用对其功能非常重要。除了它们的氧化还原配偶体，CYP已被报道与不直接参与其酶功能的其他蛋白质相互作用。在这项研究中，在小鼠肝脏中鉴定了与CYP 2C 2体内相互作用的蛋白质。Flag标记的CYP 2C 2在小鼠肝脏中外源表达，并与相关蛋白一起进行沿着的亲和纯化，所述相关蛋白通过MS鉴定并通过蛋白质印迹法确认。超过20种蛋白质可重复地与CYP 2C 2共纯化。通过蔗糖梯度离心和连续免疫沉淀分析，CYP 2C 2和相关蛋白的不均匀沉降速度与不同组成的多种CYP 2C 2复合物一致。CYP和其他药物代谢酶以及与CYP 2C 2相关的NAD/NADP需要酶的丰度表明，这些蛋白质的复合物可以提高酶促效率或促进连续的代谢步骤。伴侣蛋白可能对维持CYP 2C 2的功能很重要，而内质网蛋白是塑造内质网形态的内质网蛋白，是CYP的潜在内质网滞留蛋白，也与CYP 2C 2相关。
Interactions of microsomal cytochromes P450 (CYPs) with other proteins in the microsomal membrane are important for their function. In addition to their redox partners, CYPs have been reported to interact with other proteins not directly involved in their enzymatic function. In this study, proteins were identified that interact with CYP2C2 in vivo in mouse liver. Flag-tagged CYP2C2 was expressed exogenously in mouse liver and was affinity purified, along with associated proteins which were identified by MS and confirmed by western blotting. Over 20 proteins reproducibly co-purified with CYP2C2. The heterogeneous sedimentation velocity of CYP2C2 and associated proteins by centrifugation in sucrose gradients and sequential immunoprecipitation analysis were consistent with multiple CYP2C2 complexes of differing composition. The abundance of CYPs and other drug metabolizing enzymes and NAD/NADP requiring enzymes associated with CYP2C2 suggest that complexes of these protein may improve enzymatic efficiency or facilitate sequential metabolic steps. Chaperones, which may be important for maintaining CYP function, and reticulons, endoplasmic reticulum proteins that shape the morphology of the endoplasmic reticulum and are potential endoplasmic reticulum retention proteins for CYPs, were also associated with CYP2C2.
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