胎儿三毛滴虫质膜的分离与生化特性分析
Isolation and biochemical characterization of the plasma membrane of Tritrichomonas foetus.
作者信息
Díaz J A, Monteiro-Leal L H, de Souza W
机构信息
Laboratorio de Ultraestrutura Celular Hertha Meyer, Instituto de Biofísica Carlos Chagas Filho Universidade Federal do Rio de Janeiro, Brasil.
出版信息
Biocell. 1996 Apr;20(1):21-31.
A fraction containing plasma membrane-enriched vesicles has been prepared from Tritrichomonas foetus. Cells were ruptured using a Potter type homogenizer, under well controlled conditions, and membranes were isolated by differential centrifugation and in discontinuous sucrose gradient. This fraction was enriched 8 and 10-fold in the plasma membrane marker enzymes 5'-nucleotidase and (Na+ + K+)-dependent, ouabain-sensitive ATPase, respectively. Determination of Glucose-6-phosphatase and NADPH cytochrome c reductase activities in this fraction, indicates a minimal contamination with endoplasmic reticulum membranes. Analysis by Sodium Dodecyl Sulfate-Polyacrylamide (SDS-PAGE) gradient gel showed that the plasma membrane fraction contains several proteins with major bands corresponding to apparent molecular weights of 48, 45, 39, 37, 32, 30, 27, 23, 20, 19, 17, and 15 kDa.
已从胎儿三毛滴虫中制备出富含质膜的囊泡组分。在严格控制的条件下，使用波特型匀浆器使细胞破裂，通过差速离心和不连续蔗糖梯度分离膜。该组分中质膜标记酶5'-核苷酸酶和（Na + + K +）依赖性、哇巴因敏感的ATP酶分别富集了8倍和10倍。对该组分中葡萄糖-6-磷酸酶和NADPH细胞色素c还原酶活性的测定表明，内质网膜的污染极少。十二烷基硫酸钠-聚丙烯酰胺（SDS-PAGE）梯度凝胶分析表明，质膜组分包含几种蛋白质，主要条带对应的表观分子量为48、45、39、37、32、30、27、23、20、19、17和15 kDa。
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