ELECTROPHORETIC MAP OF ACIDIC AND NEUTRAL HUMAN SPERMATOZOAL PROTEINS

被引：16

作者：

NAABYHANSEN, S ^{[1
]}

机构：

[1] UNIV COPENHAGEN,PROT LAB,DK-1168 COPENHAGEN,DENMARK

来源：

JOURNAL OF REPRODUCTIVE IMMUNOLOGY | 1990年 / 17卷 / 03期

关键词：

2-D electroblotting; human spermatozoa; radiolabelling; surface proteins;

D O I：

10.1016/0165-0378(90)90001-M

中图分类号：

R392 [医学免疫学]; Q939.91 [免疫学];

学科分类号：

100102 ;

摘要：

A high resolution two dimensional electrophoretic technique (2-D), isoelectric focusing followed by polyacrylamide gel electrophoresis, for separation of neutral and acidic human spermatozoal proteins was established. Silver stained 2-D gels revealed over 260 proteins with Mr 20-200 kDa and pI between 4.5 and 7.8. Membrane proteins were radio-iodinated in their hydrophobic cores with the hydrophobic photoactivatable reagent 3 (trifluoromethyl)-3-(m-[125I]iodophenyl) diazirine (TID). Externally exposed carbohydrate side chains were radiolabelled by NaB(3H)4 reduction. 2-D lectin blotting was performed with horseradish peroxidase conjugated Concanavalin A (Con A), biotinylated Vicia faba agglutinin (VFA) and Solanum tuberosum agglutinin (STA). A map of 258 human spermatozoal proteins is presented and 48 TID-labelled proteins have been biochemically characterized. © 1990.

引用

页码：167 / 185

页数：19