Electrophoretic map of acidic and neutral human spermatozoal proteins.
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.
Protein Laboratorium, University of Copenhagen, Denmark.
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Naaby-Hansen
D000345:Affinity Labels; D001389:Azirines; D001894:Borohydrides; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007457:Iodine Radioisotopes; D007525:Isoelectric Focusing; D037102:Lectins / Q000378:metabolism; D008297:Male; D008562:Membrane Glycoproteins / Q000032:analysis; D008565:Membrane Proteins / Q000032:analysis*; D012995:Solubility; D013094:Spermatozoa / Q000737:chemistry*; D014316:Tritium
D000345:Affinity Labels; D001389:Azirines; D001894:Borohydrides; D007457:Iodine Radioisotopes; D037102:Lectins; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D014316:Tritium; C033174:3-(trifluoromethyl)-3-(3-iodophenyl)diazirine; C025364:sodium borohydride