小鼠肝细胞膜分级分离过程中表面抗原的分布
The distribution of surface antigens during fractionation of mouse liver plasma membranes.
作者信息
Gurd J W, Evans W H, Perkins H R
出版信息
Biochem J. 1972 Nov;130(1):271-80. doi: 10.1042/bj1300271.
Abstract
- Antiserum to purified mouse liver plasma membranes was prepared and the partially purified gamma-globulin antibody fraction was iodinated with (125)I. The reaction of the (125)I-labelled gamma-globulin antibody with isolated mouse liver plasma membranes was studied. 2. The gammaglobulin antibody bound specifically to mouse liver plasma membranes and there was little reaction with mouse liver intracellular membranes or with surface-membrane fractions from either rat liver or pig lymphocytes. 3. ;Light' and ;heavy' mouse liver plasma-membrane subfractions bound similar amounts of gamma-globulin antibody, and this is consistent with a surface origin for the light fraction. 5. Plasma membranes were fractionated by sequential extraction with 50mm-NaHCO(3)-Na(2)CO(3) buffer, pH10.2, containing 10mm-EDTA and aq. 33% (v/v) pyridine. The alkali-soluble and -insoluble fractions and the pyridine-soluble and -insoluble fractions all reacted with the antiserum, and the cross-reactivity among the various fractions and with the total plasma membranes was investigated. 5. The results are discussed in terms of the arrangement of the antigenic determinants within the membrane.
摘要
- 制备了针对纯化的小鼠肝细胞膜的抗血清，并将部分纯化的γ-球蛋白抗体组分用（125）I进行碘化。研究了（125）I标记的γ-球蛋白抗体与分离的小鼠肝细胞膜的反应。2. γ-球蛋白抗体特异性结合小鼠肝细胞膜，与小鼠肝内细胞膜或大鼠肝或猪淋巴细胞的表面膜组分几乎没有反应。3. 小鼠肝细胞膜的“轻”和“重”亚组分结合了相似量的γ-球蛋白抗体，这与轻组分起源于表面一致。5. 用含有10mM乙二胺四乙酸（EDTA）和33%（v/v）吡啶水溶液的50mM碳酸氢钠-碳酸钠缓冲液（pH10.2）顺序提取对细胞膜进行分级分离。碱溶性和碱不溶性组分以及吡啶溶性和吡啶不溶性组分均与抗血清发生反应，并研究了各组分之间以及与总细胞膜的交叉反应性。5. 根据膜内抗原决定簇的排列对结果进行了讨论。
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