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抗生物素蛋白-生物素复合物与大鼠肝细胞膜结合的部分特性

The partial characterization of the binding of avidin-biotin complex to rat liver plasma membrane.

作者信息

Chalifour L E, Dakshinamurti K

出版信息

Biochem J. 1983 Jan 15;210(1):121-8. doi: 10.1042/bj2100121.

DOI:10.1042/bj2100121
PMID:6847638
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1154197/
Abstract

Rat liver plasma membrane and rat intestinal mucosa plasma membrane were examined for their ability to bind avidin-[3H]biotin complex. Avidin-[3H]biotin complex bound specifically to liver plasma membrane with a Kd of approx. 35 nM and Bmax. of 136 pmol/mg of membrane protein. The receptor on the liver plasma membrane was exposed by Pronase, sensitive to tryptic hydrolysis and insensitive to neuraminidase. The optimum pH of specific binding was less than 6.0. Simple carbohydrates, mannan and ovalbumin did not inhibit specific binding. Periodate-treated or alpha-mannosidase-treated avidin was able to inhibit binding of either avidin-[3H]biotin complex or untreated avidin to plasma membrane. [3H]Avidin containing methylated lysine residues did not bind to rat liver plasma membrane. There was no specific binding of the avidin-biotin complex to rat intestinal mucosa plasma membrane.

摘要

检测了大鼠肝细胞膜和大鼠肠黏膜细胞膜结合抗生物素蛋白-[³H]生物素复合物的能力。抗生物素蛋白-[³H]生物素复合物特异性结合肝细胞膜，解离常数（Kd）约为35 nM，最大结合量（Bmax）为136 pmol/mg膜蛋白。肝细胞膜上的受体可被链霉蛋白酶暴露，对胰蛋白酶水解敏感，对神经氨酸酶不敏感。特异性结合的最佳pH小于6.0。单糖、甘露聚糖和卵清蛋白不抑制特异性结合。经高碘酸盐处理或α-甘露糖苷酶处理的抗生物素蛋白能够抑制抗生物素蛋白-[³H]生物素复合物或未处理的抗生物素蛋白与细胞膜的结合。含有甲基化赖氨酸残基的[³H]抗生物素蛋白不与大鼠肝细胞膜结合。抗生物素蛋白-生物素复合物与大鼠肠黏膜细胞膜无特异性结合。

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本文引用的文献

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Absence of an intracellular cobalamin-binding protein in cultured fibroblasts from patients with defective synthesis of 5'-deoxyadenosylcobalamin and methylcobalamin.5'-脱氧腺苷钴胺素和甲钴胺素合成缺陷患者培养成纤维细胞中缺乏细胞内钴胺素结合蛋白。
Proc Natl Acad Sci U S A. 1975 Nov;72(11):4617-21. doi: 10.1073/pnas.72.11.4617.
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The use of the avidin-biotin complex as a tool in molecular biology.抗生物素蛋白-生物素复合物在分子生物学中的应用。
Methods Biochem Anal. 1980;26:1-45. doi: 10.1002/9780470110461.ch1.
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The biotin requirement of HeLa cells.海拉细胞对生物素的需求。
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Low density lipoprotein binding to human platelets: role of charge and of specific amino acids.
Biochem Biophys Res Commun. 1981 Mar 16;99(1):308-18. doi: 10.1016/0006-291x(81)91746-0.
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Biochem Biophys Res Commun. 1982 Feb 11;104(3):1047-53. doi: 10.1016/0006-291x(82)91355-9.
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