{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,4]],"date-time":"2026-10-04T17:25:54Z","timestamp":1791134754296,"version":"4.1.0"},"reference-count":96,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2005,2,4]],"date-time":"2005-02-04T00:00:00Z","timestamp":1107475200000},"content-version":"vor","delay-in-days":5179,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecular Reproduction Devel"],"published-print":{"date-parts":[[1990,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>The kinetic properties and inhibitor sensitivity of human sperm phospholipase A<jats:sub>2<\/jats:sub> (PLA<jats:sub>2<\/jats:sub>; EC 3.1.1.4) were studied. Phospholipase activity was isolated from human spermatozoa by acid extraction. Hydrolysis of dipalmitoyl phosphatidylcholine was specific to the sn\u20102 position. Activity was sensitive to product inhibition (60% inhibition by 0.1 mM lysophosphatidylcholine). The effects of Ca<jats:sup>2+<\/jats:sup> and sodium deoxycholate on enzyme activity were biphasic; maximal activities were observed at 0.5 mM concentration of each agent. PLA<jats:sub>2<\/jats:sub> was stimulated (135%) by 3% dimethylsulfoxide and was inhibited by elevated ionic strength (approximately 70% inhibition with either 0.2 M NaCl or 0.2 M KCl). Two molecular forms of PLA<jats:sub>2<\/jats:sub> were kinetically distinguishable, one with an apparent Michaelis constant and maximal reaction velocity of 3.0 \u03bcM and 0.64 mlU\/mg protein and the other with respective constants of 630 \u03bcM and 32.0 mlU\/mg protein. Both forms of the enzyme were Ca<jats:sup>2+<\/jats:sup> dependent and heat stable; however, the low\u2010K<jats:sub>m<\/jats:sub> activity was less resistant to 60\u00b0C preincubation at pH 7.5 (28% inactivation of low\u2010K<jats:sub>m<\/jats:sub> activity after 45 min, as compared to no effect on high\u2010K<jats:sub>m<\/jats:sub> activity). Quinacrine was a noncompetitive PLA<jats:sub>2<\/jats:sub> inhibitor with K<jats:sub>i<\/jats:sub>s for low\u2010 and high\u2010K<jats:sub>m<\/jats:sub> activities of 0.42 mM and 0.49 mM, respectively. Trifluoperazine (calmodulin antagonist) inhibited the high\u2010K<jats:sub>m<\/jats:sub> activity noncompetitively (K<jats:sub>i<\/jats:sub> = 87 \u03bcM) and the low\u2010K<jats:sub>m<\/jats:sub> activity by a mechanism consistent with the removal of a nonessential activator. Dissociation and rate constants for inactivation of low\u2010 and high\u2010K<jats:sub>m<\/jats:sub> activities by p\u2010bromophenacyl bromide were 0.28 mM and 0.032 min<jats:sup>\u22121<\/jats:sup>, and 0.73 mM and 0.066 min<jats:sup>\u22121<\/jats:sup>, respectively. PLA<jats:sub>2<\/jats:sub> was inhibited by p\u2010nitrophenyl\u2010p\u2032\u2010guanidinobenzoate, at higher concentrations (10<jats:sup>\u22124<\/jats:sup>\u201310<jats:sup>\u22123<\/jats:sup> M) than required to inhibit trypsinlike proteinases; p\u2010aminobenzamidine, another potent trypsin\/acrosin inhibitor, stimulated (approximately 40%) PLA<jats:sub>2<\/jats:sub> at concentrations from 2\u20135 mM but inhibited PLA<jats:sub>2<\/jats:sub> (40\u201350%) at a concentration of 10 mM. MnCl<jats:sub>2<\/jats:sub> (5mM) inhibited low\u2010 and high\u2010K<jats:sub>m<\/jats:sub> PLA<jats:sub>2<\/jats:sub> activities by 77% and 76%, respectively. Quinacrine (0.4 mM), trifluoperazine (20 \u03bcM), p\u2010bromophenacyl bromide (20 \u03bcM), and MnCl<jats:sub>2<\/jats:sub> (5 mM) were tested as inhibitors of the ionophore A23187\u2010induced human acrosome reaction. Inhibition was noted only with quinacrine (32%) and MnCl<jats:sub>2<\/jats:sub> (93%). The effect of MnCl<jats:sub>2<\/jats:sub> was restricted to an interaction with A23187, rather than with PLA<jats:sub>2<\/jats:sub>; p\u2010Bromophenacyl bromide inhibited (P &lt; 0.05) PLA<jats:sub>2<\/jats:sub> (29%) when added to intact spermatozoa but had no effect on the acrosome reaction. PLA<jats:sub>2<\/jats:sub> inhibition was poorly correlated with the acrosome reaction. Results from this study suggest that (1) human sperm contain at least two kinetically distinguishable forms of PLA<jats:sub>2<\/jats:sub>, with properties similar to those of PLA<jats:sub>2<\/jats:sub> in other species and tissues; (2) PLA<jats:sub>2<\/jats:sub> inhibition is not a likely mechanism for inhibition of acrosome reactions by acrosin inhibitors; (3) sperm PLA<jats:sub>2<\/jats:sub> may be modulated by an endogenous activator; (4) the low\u2010K<jats:sub>m<\/jats:sub> PLA<jats:sub>2<\/jats:sub> is similar to the group I (pancreatic) enzyme; and (5) PLA<jats:sub>2<\/jats:sub> is not a pivotal or rate\u2010limiting step in the human acrosome reaction.<\/jats:p>","DOI":"10.1002\/mrd.1080270405","type":"journal-article","created":{"date-parts":[[2005,2,24]],"date-time":"2005-02-24T00:24:10Z","timestamp":1109204650000},"page":"305-325","source":"Crossref","is-referenced-by-count":14,"title":["Characterization and inhibitor sensitivity of human sperm phospholipase A<sub>2<\/sub>: Evidence against pivotal involvement of phospholipase A<sub>2<\/sub> in the acrosome reaction"],"prefix":"10.1002","volume":"27","author":[{"given":"Robert A.","family":"Anderson Jr.","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Susan K.","family":"Johnson","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter","family":"Bielfeld","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kenneth A.","family":"Feathergill","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lourens J. 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