Klenchin V A, Kowalchyk J A, Martin T F
Department of Biochemistry, University of Wisconsin, 433 Babcock Drive, Madison, Wisconsin, 53706, USA.
Methods. 1998 Oct;16(2):204-8. doi: 10.1006/meth.1998.0668.
A current major challenge in the study of regulated exocytosis is the identification of essential proteins that mediate the transit of secretory vesicles through trafficking stages such as recruitment, docking, and fusion. Defining the physiological roles and mechanisms of action of these essential proteins is paramount. The reconstitution of stages of regulated exocytosis in cell-free systems provides the opportunity to identify required proteins and establish their stage-specific mechanisms of action. PC12 cells, clonal cell lines of adrenal medullary origin, possess large dense-core vesicles that retain their competence for regulated exocytosis in a variety of permeable cell and isolated membrane preparations. We describe several cell-free systems for studies of regulated exocytosis derived from PC12 cells.
当前调节性胞吐作用研究中的一个主要挑战是鉴定介导分泌囊泡通过诸如募集、对接和融合等运输阶段的必需蛋白质。确定这些必需蛋白质的生理作用和作用机制至关重要。在无细胞系统中重建调节性胞吐作用阶段,为鉴定所需蛋白质并确定其阶段特异性作用机制提供了机会。PC12细胞是肾上腺髓质来源的克隆细胞系,具有大的致密核心囊泡,在各种可渗透细胞和分离膜制剂中保持其调节性胞吐作用的能力。我们描述了几种源自PC12细胞的用于研究调节性胞吐作用的无细胞系统。