Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.2.1.36 (hyaluronidase)
 4,606 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Two methods for the extraction of acrosomal membranes and enzymes from both human and rabbit spermatozoa were compared. Treatment of spermatozoa with hypotonic MgCl2 (0.05 M) solution causes removal of the plasma membrane, vesiculation, disruption and removal of the outer acrosomal membrane posterior to the equatorial segment with accompanying loss of soluble acrosomal material. Subsequent exposure to Hyamine 2389 and Triton X-100 removes acrosomal material bound to the inner acrosomal membrane with concomitant solubilization of this membrane. The MgCl2 extract from rabbit spermatozoa contained a higher yield of hyaluronidase, acrosin, and total proteinase activities, whereas the subsequent detergent extracts contained higher yields of both arylsulfatase A and B activities. By comparison, after 4 minutes of sonication to separate heads and tails, both rabbit and human spermatozoa when viewed by transmission electron microscopy showed alterations of plasma and outer acrosomal membranes with considerable loss of the acrosomal contents. Analysis of acrosomal enzymes indicates the greatest percentage of all the enzymes assayed was located in the extract obtained by sonication in contrast to either the separated head or tail fractions used for further subcellular extraction. Subsequent treatment with Hyamine and Triton yields only minimal amounts of enzyme activity.
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PMID:Extraction of human and rabbit acrosomes: a comparison of sequential and sonication methods. 51 71

Arylsulfatase A was extracted and purified from boar epididymal sperm acrosomes. Acrosomes were extracted by sonication in 50 mM Tris-maleate buffer containing 50 mM MgCl2, pH 6.1, followed by treatment with 50 mM Tris-maleate plus 0.2% Brij-35, pH 6.1. Purification of arylsulfatase A was performed with a three-step procedure consisting of centrifugation (85,000 X g), affinity chromatography with p-aminobenzamidine-Sepharose followed by chromatography on diethyaminoethyl (DEAE) Sephadex. The specific activity of the purified enzyme was 54 mumol/h per mg protein. The purified arylsulfatase did not contain any detectable acrosin or hyaluronidase activities. Sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis revealed a major band with an estimated molecular weight of 65,000 daltons. Properties of arylsulfatase A, determined by hydrolysis of p-nitrocatechol sulfate, indicated that the enzyme was inhibited 46% by 3.1 microM Ag+ and had a pH optimum of 4.2. Boar acrosomal arylsulfatase A dispersed the cumulus cells of ovulated hamster and rabbit eggs as well as those of follicular pig eggs. No effect of the enzyme on the zona pellucida or the oolemma was observed.
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PMID:Purification of boar acrosomal arylsulfatase A and possible role in the penetration of cumulus cells. 614 55

Cumulus cell layers of expanded cumulus oocyte complexes (COCs) are interlinked with networks of hyaluronic acid, chondroitin sulfate B proteoglycans and link proteins, and they can be dispersed by sperm surface hyaluronidases. In this report, we showed that arylsulfatase A (AS-A), existing on the sperm head surface, also had this dispersion action. Purified AS-A free of protease, hyaluronidase and chondroitinase activities could disperse the cumulus matrix of expanded COCs. However, this COC dispersion action was not associated with AS-A desulfation activity, assayed by using p-nitrocatecholsulfate (artificial substrate). COCs incubated for 1 h with sperm pretreated with anti-AS-A IgG in the presence of apigenin (a hyaluronidase inhibitor) did not exhibit matrix dispersion, whereas several cumulus layers were already dispersed in COCs incubated with sperm pretreated with preimmune IgG. Furthermore, sperm from AS-A null mice showed a significant delay in COC dispersion, compared with wild-type sperm. Within 1 h of sperm-COC co-incubation, the size of COCs incubated with AS-A null sperm was 65% of the original dimension, whereas that of COCs inseminated with wild-type sperm was only 17%. A further delay in COC dispersion by AS-A(-/-) mouse sperm was observed when apigenin was present in the co-incubation. We also showed for the first time that AS-A had a specific affinity for chondroitin sulfate B, a component of cumulus matrix proteoglycan networks; this might provide a mechanism of cumulus matrix destabilization induced by sperm surface AS-A.
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PMID:Sperm surface arylsulfatase A can disperse the cumulus matrix of cumulus oocyte complexes. 1747 85