从暹罗鳄中纯化和鉴定卵转铁蛋白。

Purification and characterization of ovotransferrin from Crocodylus siamensis.

机构信息

Department of Biochemistry, Faculty of Science, Khon Kaen University, Khon Kaen 40002, Thailand.

出版信息

Protein J. 2013 Feb;32(2):89-96. doi: 10.1007/s10930-012-9461-2.

Abstract

Ovotransferrin (OTf) is the major glycoprotein in reptile egg whites. However, knowledge concerning its functional and biological properties remains limited. In this study, OTf from Crocodylus siamensis was purified and characterized. The proteins were precipitated with 80 % ammonium sulfate and then purified by anion exchange chromatography followed by hydrophobic interaction chromatography. The purified crocodile ovotransferrin (cOTf) had a molecular weight of 79 kDa. Analysis by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) indicated multiple isoforms of cOTf, which had isoelectric points ranging from 6.0 to 6.8. cOTf was N-linked glycosylated protein identified by using PNGase F deglycosylation technique. Optimal autoproteolysis of cOTf occurred under acidic conditions and pH values more than 5, which differs from that of OTf.

摘要

卵转铁蛋白（OTf）是爬行动物蛋清中的主要糖蛋白。然而，其功能和生物学特性的相关知识仍然有限。在这项研究中，从暹罗鳄中纯化和表征了 OTf。使用 80%的硫酸铵沉淀蛋白质，然后通过阴离子交换色谱和疏水相互作用色谱进行纯化。纯化的鳄卵转铁蛋白（cOTf）的分子量为 79 kDa。二维聚丙烯酰胺凝胶电泳（2D-PAGE）分析表明，cOTf 具有多种同工型，等电点范围为 6.0 至 6.8。通过使用 PNGase F 去糖基化技术鉴定，cOTf 是一种 N-连接的糖蛋白。cOTf 的最佳自切发生在酸性条件下和 pH 值大于 5 的条件下，这与 OTf 不同。