Phospholipases A2 in the reproductive system of the bull.
1. Phospholipase A2 activities were studied in the reproductive organs, seminal plasma and spermatozoa of adult bulls. 2. Phosphatidylethanolamine and phosphatidylcholine with 14C-labelled linoleic (lino-PE, lino-PC) or arachidonic acid (ara-PE, ara-PC) at sn-2 position as well as a fluorescent derivative (4-pyrenylbutyric acid) of phosphatidylcholine (PPC) were used as substrates. 3. The radioactive substrates were hydrolysed most strongly by homogenates of the prostate and Cowper's gland, but also seminal vesicle and its secretory fluid, seminal plasma and ejaculated spermatozoa contained hydrolytic activity. The fluorescence substrate was most strongly hydrolysed by homogenates of ampulla and seminal vesicle as well as its secretory fluid, seminal plasma and ejaculated spermatozoa. 4. Seminal plasma and seminal vesicle fluid contained a Ca2(+)-independent enzyme (enzyme I), which hydrolysed only PPC, while another Ca2(+)-dependent enzyme (enzyme II) hydrolysed only the radioactive substrates. 5. Both enzymes were purified from the seminal vesicle fluid and their biochemical properties were analysed. In SDS-PAGE enzyme I preparation resulted in two major bands with molecular weights of 16,000 and 60,000 in equal quantities and minor band at 15,000. The binding of the enzyme I to Con A-Sepharose indicated that it is a glycoprotein and it had multiple pI-values from 3.75 to 5.0. Enzyme II gave in SDS-PAGE two closely located bands with molecular weights of about 15,000 and 16,000 (major band). Isoelectric focusing showed one band at pI 4.7. Both enzymes appear to bind to spermatozoa at ejaculation but their function remains to be shown.
Rönkkö
S
S
Department of Anatomy, University of Kuopio, Finland.
Lahtinen
R
R
Vanha-Perttula
T
T
D000818:Animals; D002030:Bulbourethral Glands / Q000201:enzymology; D002417:Cattle; D002845:Chromatography; D005837:Genitalia, Male / Q000201:enzymology*; D006868:Hydrolysis; D007700:Kinetics; D008297:Male; D009928:Organ Specificity; D010741:Phospholipases A / Q000302:isolation & purification / Q000378:metabolism*; D054467:Phospholipases A2; D011467:Prostate / Q000201:enzymology; D012661:Semen / Q000201:enzymology*; D012669:Seminal Vesicles / Q000201:enzymology; D013094:Spermatozoa / Q000201:enzymology*; D013379:Substrate Specificity
D010741:Phospholipases A; D054467:Phospholipases A2