Two satisfactory methods for purification of human acrosin.
Acrosin has been purified from human sperm cells by two alternative procedures which give purer products and in higher yields than could be achieved previously. The products were characterized by their molecular weight, catalytic action, sensitivity to inhibitors, and reaction with a polyclonal anti-acrosin antibody. After acid extraction of the cells, one method involves removal of acrosin inhibitors by vacuum dialysis, followed by affinity chromatography on a soybean trypsin inhibitor (SBTI) column, and therefore requires that the acrosin be in an active form capable of binding to the inhibitor. The other method involves affinity chromatography on a column of a monoclonal anti-acrosin antibody (MAb) and can be used to provide either active or proenzyme forms of acrosin, by choice of extraction conditions and inclusion of appropriate inhibitors. The yield of human acrosin from the SBTI method was 104% and from the MAb column was 75%. It is hoped that these procedures will make the very scarce human acrosin more readily available for further study.
D000176:Acrosin / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D000911:Antibodies, Monoclonal; D002846:Chromatography, Affinity; D010450:Endopeptidases / Q000302:isolation & purification*; D006801:Humans; D007700:Kinetics; D008297:Male; D012661:Semen / Q000201:enzymology*
D000911:Antibodies, Monoclonal; D010450:Endopeptidases; D000176:Acrosin