Bissig Christin, Johnson Shem, Gruenberg Jean
Biochemistry Department, University of Geneva, 30 quai Ernest Ansermet, 1211 Geneva 4, Switzerland.
Methods Cell Biol. 2012;108:19-46. doi: 10.1016/B978-0-12-386487-1.00002-X.
Endosomes along the degradation pathway exhibit a multivesicular appearance and differ in their lipid compositions. Association of proteins to specific membrane lipids and presumably also lipid-lipid interactions contribute to the formation of functional membrane platforms that regulate endosome biogenesis and function. This chapter provides a brief review of the functions of endosomal lipids in the degradation pathway, a discussion of techniques that allow studying lipid-based mechanisms and a selection of step-by-step protocols for in vivo and in vitro methods commonly used to study lipid roles in endocytosis. The techniques described here have been used to elucidate the function of the late endosomal lipid lysobisphosphatidic acid and allow the monitoring of lipid distribution, levels and dynamics, as well as the characterization of lipid-binding partners.
沿降解途径的内体呈现多囊泡外观，且脂质组成不同。蛋白质与特定膜脂的结合以及可能的脂-脂相互作用有助于形成调节内体生物发生和功能的功能性膜平台。本章简要综述了内体脂质在降解途径中的功能，讨论了用于研究基于脂质机制的技术，并挑选了常用于研究脂质在胞吞作用中作用的体内和体外方法的分步方案。此处描述的技术已用于阐明晚期内体脂质溶血双磷脂酸的功能，并可监测脂质分布、水平和动态，以及鉴定脂质结合伴侣。
