Co-immunoprecipitation for identifying protein-protein interaction on lipid droplets.

Xiaochuan Fu, Shuyan Zhang, Pingsheng Liu
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引用次数: 0

Abstract

The lipid droplet (LD) is a conserved organelle that exists in almost all organisms, ranging from bacteria to mammals. Dysfunctions in LDs are linked to a range of human metabolic syndromes. The formation of protein complexes on LDs is crucial for maintaining their function. Investigating how proteins interact on LDs is essential for understanding the role of LDs. We have developed an effective method to uncover protein-protein interactions and protein complexes specifically on LDs. In this method, we conduct co-immunoprecipitation (co-IP) experiments using LD proteins extracted directly from isolated LDs, rather than utilizing proteins from cell lysates. To elaborate, we begin by purifying LDs with high-quality and extracting LD-associated proteins. Subsequently, the co-IP experiment is performed on these LD-associated proteins directly, which would enhance the co-IP experiment specificity of LD-associated proteins. This method enables researchers to directly unveil protein complexes on LDs and gain deeper insights into the functional roles of proteins associated with LDs.

共免疫沉淀法用于识别脂滴上蛋白质与蛋白质之间的相互作用。
脂滴(LD)是一种保守的细胞器,存在于从细菌到哺乳动物等几乎所有生物体内。脂滴的功能障碍与一系列人类代谢综合征有关。LD 上蛋白质复合物的形成对维持其功能至关重要。研究蛋白质如何在 LDs 上相互作用对于了解 LDs 的作用至关重要。我们已经开发出一种有效的方法,专门用于揭示 LDs 上的蛋白质-蛋白质相互作用和蛋白质复合物。在这种方法中,我们使用直接从分离的 LDs 中提取的 LD 蛋白进行共沉淀(co-immunoprecipitation,co-IP)实验,而不是利用细胞裂解液中的蛋白质。具体来说,我们首先用高质量的方法纯化 LD,并提取 LD 相关蛋白。随后,直接对这些 LD 相关蛋白进行 co-IP 实验,这将提高 LD 相关蛋白 co-IP 实验的特异性。这种方法能让研究人员直接揭示 LD 上的蛋白质复合物,并深入了解与 LD 相关的蛋白质的功能作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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