Identification and partial characterization of plasma membrane polypeptides of Trypanosoma brucei.
Identification and partial characterization of plasma membrane polypeptides of Trypanosoma brucei.
复制标题
布氏锥虫质膜多肽的鉴定和部分表征。
DOI：
10.1016/0005-2736(82)90351-0
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发表时间：
1982
期刊：
影响因子：
--
通讯作者：
Curtis L. Patton
中科院分区：
文献类型：
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作者：
Patricia E. Mancini;James E. Strickler;Curtis L. Patton
用超声处理和蔗糖梯度差速离心法从布氏锥虫中制备了一种富含质膜的囊泡组分。该级分在质膜标记酶腺苷酸环化酶（EC 4.6）中富集5倍。1.1)和哇巴因可降解的（Na++ K+）依赖性腺苷三磷酸酶（EC 3.6. 1.3）。它还富含碘化表面蛋白高达14倍，[3 H]甘露糖标记的糖蛋白高达4倍，其中主要的可变表面外壳糖蛋白是主要成分。质膜组分和其他亚细胞组分的蛋白质已通过十二烷基硫酸钠-聚丙烯酰胺梯度平板凝胶电泳分析鉴定。几个高分子量的表面糖肽已被选择性地研究和部分特征的结合代谢标记与[3 H]甘露糖，乳过氧化物酶催化的表面碘化，和亲和色谱上的Con A-琼脂糖。除了主要的可变表面包衣糖蛋白（估计Mr = 58 000），有几个次要的表面糖肽（Mr = 76 000，86 000和92 000-100 000），这是明显的外源性膜成分，和两个表面糖肽（Mr = 42 000和130 000），这是内在的膜成分。
A plasma membrane-enriched vesicle fraction has been prepared from Trypanosoma brucei by sonication and differential centrifugation on sucrose gradients. This fraction is enriched 5-fold in the plasma membrane marker enzymes adenyl cyclase (EC 4.6. 1.1) and ouabain-inhibitable,(Na++ K+)-dependent adenosine triphosphatase (EC 3.6. 1.3). It is also enriched up to 14-fold in iodinated surface proteins, and up to 4-fold in [3 H] mannose-labeled glycoproteins, of which the major variable surface coat glycoprotein is the main constituent. Proteins of the plasma membrane fraction and other subcellular fractions have been identified by electrophoretic analysis in sodium dodecyl sulfate-polyacrylamide gradient slab gels. Several high molecular weight surface glycopeptides have been selectively investigated and partially characterized by a combination of metabolic labeling with [3 H] mannose, lactoperoxidase-catalyzed surface iodination, and affinity chromatography on Con A-Sepharose. In addition to the major variable surface coat glycoprotein (estimated M r= 58 000), there are several minor surface glycopeptides (M r= 76 000, 86 000 and 92 000–100 000) which are apparent extrinsic membrane components, and two surface glycopeptides (M r= 42 000 and 130 000) which are intrinsic membrane components.