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
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
收藏
页码:167 / 185
页数:19
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