Isolation and characterization of a protein with homology to angiotensin converting enzyme from the periacrosomal plasma membrane of equine spermatozoa.
Abstract: The periacrosomal plasma membrane of spermatozoa is involved in sperm binding to oviductal epithelial cells and to the zona pellucida. A protein of 68-70 kD molecular mass was purified biochemically from the isolated periacrosomal plasma membrane of equine spermatozoa as a possible receptor for adhesion of spermatozoa to oviductal epithelial cells. A polyclonal antibody raised in rabbits against the purified equine sperm membrane protein recognized the 70 kD and an antigenically related to 32 kD protein in preparations of isolated periacrosomal sperm plasma membrane and in detergent extracted ejaculated and epididymal spermatozoa. A larger protein (approximately 110 kD) was detected in equine testis. Two antigenically related proteins (64 and 45 kD) were recognized on the plasma membrane of cynomolgus macaque spermatozoa. In vitro sperm-binding assays were performed in the presence of antigen-binding fragments or IgG purified from the polyclonal antiserum to investigate a possible function to the isolated protein in binding of equine spermatozoa to homologous oviductal epithelial cells or zona pellucida. Incubation with antigen-binding fragments or IgG purified from the antiserum did not inhibit binding of equine spermatozoa either to oviductal epithelial cells or the zona pellucida. On ultrastructural examination, the antibody bound exclusively to the cytoplasmic side of the periacrosomal plasma membrane of equine and macaque spermatozoa. Microsequence analysis of 13 residues of sequence showed strong homology with a number of angiotensin converting enzymes: An 84% identity was identified with testis specific and somatic forms of human and mouse angiotensin-converting enzyme. Immunocytochemistry and immunoblot analysis established that the protein is specific for the periacrosomal membrane of ejaculated, epididymal, and testicular stallion spermatozoa.
Publication Date: 1997-09-18 PubMed ID: 9291475DOI: 10.1002/(SICI)1098-2795(199710)48:2<251::AID-MRD13>3.0.CO;2-0Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research identifies and examines a protein comparable to angiotensin converting enzyme in the periacrosomal plasma membrane of horse sperm. The study investigates this protein’s role in sperm binding to female reproductive cells and presents the protein’s characteristics.
Identification and Isolation of the Protein
- A protein of 68-70 kiloDaltons (kD) was biochemically purified from the periacrosomal plasma membrane of equine spermatozoa. The purification was part of an attempt to identify a potential receptor for sperm adhesion to oviductal epithelial cells.
- An antibody made in rabbits against the purified protein recognized it, and a related protein, in preparations of isolated periacrosomal sperm plasma membrane and in detergent washed ejaculated and epididymal sperm.
- A larger approximate 110 kD protein was found in horse testis, and two antigenically comparable proteins of 64 and 45 kD were identified on the plasma membrane of cynomolgus macaque spermatozoa.
Protein Function Investigation
- Experiments were carried out in the presence of antigen-binding fragments or IgG from the polyclonal antiserum to understand this isolated protein’s function in equine sperm binding to oviductal epithelial cells or zona pellucida, a layer surrounding the egg cell.
- However, neither antigen-binding fragments nor the IgG from the antiserum inhibited the binding of horse sperm to either oviductal epithelial cells or the zona pellucida.
Structural Analysis and Homology
- On a more detailed level, the sponge-like antibody bound exclusively to the cytoplasmic side of the periacrosomal plasma membrane of horse and macaque sperm.
- The microsequence analysis of 13 residues revealed strong homology (84% identity) with other angiotensin converting enzymes, including human and mouse-specific and somatic forms.
- Using immunocytochemistry and immunoblot analysis, researchers confirmed that the protein is specifically found on the periacrosomal membrane of ejaculated, epididymal, and testicular stallion sperm.
Cite This Article
APA
Dobrinski I, Ignotz GG, Fagnan MS, Yudin SI, Ball BA.
(1997).
Isolation and characterization of a protein with homology to angiotensin converting enzyme from the periacrosomal plasma membrane of equine spermatozoa.
Mol Reprod Dev, 48(2), 251-260.
https://doi.org/10.1002/(SICI)1098-2795(199710)48:2<251::AID-MRD13>3.0.CO;2-0 Publication
Researcher Affiliations
- Department of Clinical Sciences, Cornell University, Ithaca, New York, USA.
MeSH Terms
- Acrosome / enzymology
- Animals
- Cell Membrane / chemistry
- Epididymis
- Fallopian Tubes / metabolism
- Female
- Horses
- Humans
- Immunosorbent Techniques
- Male
- Mice
- Microscopy, Electron
- Molecular Weight
- Peptide Mapping
- Peptidyl-Dipeptidase A / chemistry
- Proteins / chemistry
- Proteins / isolation & purification
- Rabbits
- Spermatozoa / metabolism
- Spermatozoa / ultrastructure
- Testis
Citations
This article has been cited 3 times.- Gianzo M, Subirán N. Regulation of Male Fertility by the Renin-Angiotensin System. Int J Mol Sci 2020 Oct 26;21(21).
- Kerns K, Zigo M, Sutovsky P. Zinc: A Necessary Ion for Mammalian Sperm Fertilization Competency. Int J Mol Sci 2018 Dec 18;19(12).
- Hagaman JR, Moyer JS, Bachman ES, Sibony M, Magyar PL, Welch JE, Smithies O, Krege JH, O'Brien DA. Angiotensin-converting enzyme and male fertility. Proc Natl Acad Sci U S A 1998 Mar 3;95(5):2552-7.
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