Detection of prophospholipase A2 in human spermatozoa.
Prophospholipase A2 (proPA2) has been isolated from human spermatozoa after acid extraction and chromatography on hydrophobic WP-Butyl (C4) and ion-exchange (SP 5PW) columns. The addition of benzamidine, a noncompetitive synthetic trypsin inhibitor, to semen samples has kept a portion of the sperm phospholipase A2 (PA2) in its zymogen form and allowed its isolation after acid extraction. When radioactive phosphatidylcholine (PC) or phosphatidylethanolamine (PE) were used as substrates, an identical elution profile of this enzyme was obtained on a C4 column. The proenzyme was separated from active PA2 on the C4 column. Human sperm proPA2 exhibited a less cationic charge than active PA2 on the SP 5PW column. Porcine pancreatic proPA2 had the same chromatographic behavior on high performance liquid chromatography (HPLC) (SP 5PW) as human sperm proPA2. The purification procedure resulted in the isolation of proPA2 which, upon activation by proteolysis, presented the same chromatographic elution profile on HPLC as active PA2 of human spermatozoa and porcine pancreas. Thus, a zymogen form of PA2 exists in human spermatozoa.
Antaki
P
P
Department of Biochemistry and of Medicine University of Montreal Maisonneuve-Rosemont Hospital Research Center, Quebec, Canada.
Guérette
P
P
Chapdelaine
A
A
Roberts
K D
KD
D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004792:Enzyme Precursors / Q000032:analysis / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D010740:Phospholipases / Q000302:isolation & purification*; D010741:Phospholipases A / Q000032:analysis / Q000302:isolation & purification*; D054467:Phospholipases A2; D013094:Spermatozoa / Q000201:enzymology*
D004792:Enzyme Precursors; D010740:Phospholipases; D010741:Phospholipases A; D054467:Phospholipases A2