METHODS FOR SEPARATION AND DEGLYCOSYLATION OF MUCIN SUBUNITS

被引:68
作者
THORNTON, DJ [1 ]
HOWARD, M [1 ]
DEVINE, PL [1 ]
SHEEHAN, JK [1 ]
机构
[1] UNIV QUEENSLAND,DEPT OBSTET & GYNAECOL,HERSTON,QLD 4029,AUSTRALIA
基金
英国惠康基金;
关键词
D O I
10.1006/abio.1995.1266
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Mucins (density 1.35-1.50 g/ml) prepared from human cervical, gastric, respiratory, and salivary secretions were reduced and alkylated to generate reduced mucin subunits, These subunits were separated into different populations analytically by using agarose gel electrophoresis and preparatively by using anion-exchange chromatography. Both techniques separate the molecules primarily on the basis of their inherent charge. A procedure has been developed for the efficient deglycosylation of small amounts of these reduced mucin subunits immobilized on polyvinylidene difluoride. The combination of these procedures has allowed the isolation and identification of glycoforms of a specific mucin gene product in a respiratory mucous secretion. (C) 1995 Academic Press, Inc.
引用
收藏
页码:162 / 167
页数:6
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