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人κ-酪蛋白的分离与特性

Isolation and properties of human kappa-casein.

作者信息

Yamauchi K, Azuma N, Kobayashi H, Kaminogawa S

出版信息

J Biochem. 1981 Oct;90(4):1005-12. doi: 10.1093/oxfordjournals.jbchem.a133552.

Abstract

Human kappa-casein was isolated from human whole casein by gel filtration with Sephadex G-200 and hydroxylapatite chromatography in the presence of sodium dodecyl sulfate (SDS). The kappa-casein was calcium-insensitive and did stabilize human beta-casein and bovine alpha s1-casein against precipitation by calcium ions. Formation of micelles from human beta- and kappa-caseins, and calcium ions was confirmed by electron microscopic observation. On SDS-polyacrylamide gel electrophoresis (SDS-PAGE), a single band was obtained. The formation of para-kappa-caseins by chymosin was confirmed by SDS-PAGE. Two para-kappa-caseins with apparent molecular weights of 13,000 and 11,000 appeared. The molecular weight of intact human kappa-casein was estimated to be approximately 33,000. The human kappa-casein contained about 40% carbohydrate (15% galactose, 3% fucose, 15% hexosamines, and 5% sialic acid) and 0.10% (1 mol/mol) phosphorus. Its amino acid composition was similar to that of bovine kappa-casein except for serine, glutamic acid, and lysine contents.

摘要

通过在十二烷基硫酸钠(SDS)存在的情况下,使用葡聚糖凝胶G - 200进行凝胶过滤和羟基磷灰石色谱法,从人全酪蛋白中分离出人κ-酪蛋白。κ-酪蛋白对钙不敏感,并且确实能稳定人β-酪蛋白和牛αs1-酪蛋白,使其不被钙离子沉淀。通过电子显微镜观察证实了人β-酪蛋白、κ-酪蛋白和钙离子形成了胶束。在SDS - 聚丙烯酰胺凝胶电泳(SDS - PAGE)中,得到了一条单一的条带。通过SDS - PAGE证实了凝乳酶形成了副κ-酪蛋白。出现了两种表观分子量分别为13,000和11,000的副κ-酪蛋白。完整的人κ-酪蛋白的分子量估计约为33,000。人κ-酪蛋白含有约40%的碳水化合物(15%半乳糖、3%岩藻糖、15%己糖胺和5%唾液酸)和0.10%(1摩尔/摩尔)的磷。除了丝氨酸、谷氨酸和赖氨酸的含量外,其氨基酸组成与牛κ-酪蛋白相似。

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