Chicken acrosin: extraction and purification.
The objective of the present research was to identify a procedure whereby chicken acrosin could be purified. Acrosin, as evidenced by amidase activity, was extracted with urea most efficiently at a concentration of 6 M. Extraction efficiency was enhanced by spermatozoal lysis prior to admixture with 6 M urea. Lysis was induced by passage of spermatozoal suspensions through a French pressure cell. Acrosin was purified by using gel filtration, chromatofocusing, and affinity chromatography. Based on amidase activity, a 19-fold purification was obtained with a 28% recovery. Native electrophoresis resolved two major protein bands with proteolytic activity. The methods described afford the procurement of milligram amounts of chicken acrosin.
Froman
D P
DP
Department of Poultry Science, Oregon State University, Corvallis 97331.
D000176:Acrosin / Q000302:isolation & purification* / Q000378:metabolism; D000581:Amidohydrolases / Q000378:metabolism; D000818:Animals; D002645:Chickens / Q000378:metabolism*; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D008297:Male; D012661:Semen / Q000201:enzymology*; D012697:Serine Endopeptidases / Q000302:isolation & purification*; D012995:Solubility; D014508:Urea / Q000378:metabolism
D014508:Urea; D012697:Serine Endopeptidases; D000176:Acrosin; D000581:Amidohydrolases