一种用于测量组织中微管蛋白聚合和解聚形式的灵敏方法。
A sensitive method for measuring polymerized and depolymerized forms of tubulin in tissues.
作者信息
Pipeleers D G, Pipeleers-Marichal M A, Sherline P, Kipnis D M
出版信息
J Cell Biol. 1977 Aug;74(2):341-50. doi: 10.1083/jcb.74.2.341.
Abstract
A rapid method for measuring polymerized and depolymerized forms of tubulin in tissues has been developed. The procedure consists of homogenization and centrifugation of the tissue in a microtubule-stabilizing solution and depolymerization of the precipitated microtubules; polymerized and depolymerized forms of tubulin are quantitated by a colchicine-binding assay. The validity of the technique was assessed by electron microscopy and recovery studies with labeled and unlabeled preparations of polymerized and depolymerized forms of rat brain tubulin. The sensitivity of this technique allows quantitation of tubulin in 150 micrograms of tissue, wet weight. The method demonstrated that both the polymerized and depolymerized forms of tubulin were present in rat liver cells, and that in the fed state 31.3 +/-0.7% of the total tubulin pool was in the polymerized form.
摘要
已开发出一种用于测量组织中微管蛋白聚合态和解聚态的快速方法。该程序包括在微管稳定溶液中对组织进行匀浆和离心，以及沉淀微管的解聚；通过秋水仙碱结合测定法定量微管蛋白的聚合态和解聚态。通过电子显微镜以及对大鼠脑微管蛋白聚合态和解聚态的标记和未标记制剂进行回收研究，评估了该技术的有效性。该技术的灵敏度允许对湿重150微克组织中的微管蛋白进行定量。该方法表明，大鼠肝细胞中同时存在微管蛋白的聚合态和解聚态，并且在进食状态下，总微管蛋白库的31.3±0.7%处于聚合态。
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