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克氏原螯虾胃石中一种不溶性基质蛋白的增溶与化学特性分析

Solubilization and chemical characterization of an insoluble matrix protein in the gastroliths of a crayfish, Procambarus clarkii.

作者信息

Ishii K, Tsutsui N, Watanabe T, Yanagisawa T, Nagasawa H

机构信息

Ocean Research Institute, University of Tokyo, Japan.

出版信息

Biosci Biotechnol Biochem. 1998 Feb;62(2):291-6. doi: 10.1271/bbb.62.291.

Abstract

The gastrolith of the crayfish Procambarus clarkii contains a small amount of an organic matrix that is mainly chitin and proteins, together with a large amount of calcium carbonate. As the first step to understand the mechanism of calcification, we tried to characterize matrix proteins in the gastrolith. An insoluble matrix protein, referred to as gastrolith matrix protein, was made soluble with 1% SDS containing 10 mM dithiothreitol, and was purified by reverse-phase high-performance liquid chromatography. The protein had a molecular weight of about 50,500 and a blocked amino terminus. By enzymatic digestion and microsequencing, five partial amino acid sequences with a total of 225 amino acid residues were identified and found to include a repetitive sequence not reported previously.

摘要

克氏原螯虾的胃石含有少量主要由几丁质和蛋白质组成的有机基质,以及大量碳酸钙。作为了解钙化机制的第一步,我们试图对胃石中的基质蛋白进行表征。一种不溶性基质蛋白,即胃石基质蛋白,用含有10 mM二硫苏糖醇的1%十二烷基硫酸钠使其溶解,并通过反相高效液相色谱法进行纯化。该蛋白分子量约为50,500,氨基末端封闭。通过酶切和微量测序,鉴定出五个共225个氨基酸残基的部分氨基酸序列,发现其中包含一个此前未报道的重复序列。

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