PROTEINS ASSOCIATED W/ CAVEOLAE FROM ADIPOCYTES

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 脂肪细胞在胰岛素依赖性调节有机体燃料代谢中起重要作用，并以与任何其他细胞类型一样高或更高的水平表达小窝。最近，胰岛素信号传导和小窝之间的联系已被提出。然而，脂肪细胞小窝一直是相对较少的研究的主题，其内容已被最低限度的特点。借助一种新的单克隆抗体，我们开发了一种快速的程序，免疫分离的小窝来自质膜的脂肪细胞，我们的特点是它们的蛋白质含量。我们发现，免疫纯化的脂肪细胞小窝有一个相对有限的蛋白质组成，他们缺乏筏蛋白，flotillin和胰岛素受体。免疫金标记和脂肪细胞质膜的电子显微镜证实了小窝中缺乏胰岛素受体。除了小窝蛋白，小窝的结构组分，它们的主要蛋白质成分，是氨基脲敏感的胺氧化酶和清道夫脂蛋白受体CD 36。正在进行的研究，采用质谱分析的重点是确定额外的蛋白质成分和脂质补体与小窝。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Adipocytes play an important role in the insulin-dependent regulation of organismal fuel metabolism and express caveolae at levels as high or higher than any other cell type. Recently, a link between insulin signaling and caveolae has been suggested. Nevertheless, adipocyte caveolae have been the subject of relatively few studies, and their contents have been minimally characterized. With the aid of a new monoclonal antibody, we developed a rapid procedure for the immunoisolation of caveolae derived from the plasma membrane of adipocytes, and we characterized their protein content. We find that immunopurified adipocyte caveolae have a relatively limited protein composition, and they lack the raft protein, flotillin, and insulin receptors. Immunogold labeling and electron microscopy of the adipocyte plasma membrane confirmed the lack of insulin receptors in caveolae. In addition to caveolins, the structural components of caveolae, their major protein constituents, are the semicarbazide-sensitive amine oxidase and the scavenger lipoprotein receptor CD36. Ongoing studies that employ mass spectral analysis focus on identifying additional protein components and the lipid complement associated with caveolae.