犬α-1-抗蛋白酶的纯化与特性分析

Purification and characterization of canine alpha-1-antiproteinase.

作者信息

Abrams W R, Kimbel P, Weinbaum G

出版信息

Biochemistry. 1978 Aug 22;17(17):3556-61. doi: 10.1021/bi00610a021.

Abstract

The principal canine plasma protease inhibitor, alpha-1-antiproteinase, has been purified 90-fold with a 25% yield to apparent homogeneity. The purification scheme includes anion-exchange chromatography, to separate away the bulk of the serum albumin; affinity chromatography by insolubilized concanavalin A, to remove most of the other serum proteins as well as traces of albumin; and, finally, sizing on Sephacryl-S-200. Unique to this purification scheme is the batch use of insolubilized hemoglobin--Sepharose beads to remove the ubiquitous contaminant haptoglobin. The purified material has an apparent molecular weight of 58 000, 11.2% carbohydrate, and an E280nm1% = 5.82, and can be separated by isoelectric focusing into at least two distinct forms with pI values of 4.40 and 4.52. In addition, canine alpha-1-antiproteinase is immunologically distinct from human alpha-1-antiproteinase.

摘要

犬血浆主要蛋白酶抑制剂α1抗蛋白酶已被纯化90倍，产率为25%，达到表观均一性。纯化方案包括阴离子交换色谱法，以分离大部分血清白蛋白；用固定化伴刀豆球蛋白A进行亲和色谱法，以去除大部分其他血清蛋白以及痕量白蛋白；最后，在Sephacryl-S-200上进行分子筛分离。该纯化方案的独特之处在于批量使用固定化血红蛋白-琼脂糖珠以去除普遍存在的污染物触珠蛋白。纯化后的物质表观分子量为58000，碳水化合物含量为11.2%，E280nm1% = 5.82，通过等电聚焦可分离为至少两种不同形式，其等电点值分别为4.40和4.52。此外，犬α1抗蛋白酶在免疫学上与人α1抗蛋白酶不同。