Characterization of human sperm protein recognized by monoclonal antibody HS-8

0380394 - BTÚ 2013 RIV DK eng A - Abstrakt
Margaryan, Hasmik - Čapková, Jana - Pěknicová, Jana
Characterization of human sperm protein recognized by monoclonal antibody HS-8.
American Journal of Reproductive Immunology. Roč. 67, Issue Supplement s1 (2012), s. 27-28. ISSN 1046-7408. E-ISSN 1600-0897.
[13th International Symposium for Immunology of reproduction "From the roots to the tops of Reproductive Immunology". 22.06.2012-24.06.2012, Varna]
Grant CEP: GA ČR(CZ) GA523/09/1793; GA ČR(CZ) GAP503/12/1834
Výzkumný záměr: CEZ:AV0Z50520701
Klíčová slova: monoclonal antibody * GAPDHS * human sperm proteins * mass spectrometry
Kód oboru RIV: EC - Imunologie

Problem: Characterization of the sperm protein detected by monoclonal antibody (MoAb) Hs-8, which was prepared in our laboratory by immunization of BALB/c mice with human ejaculated sperms. Method of Study: Indirect immunofluorescence, electrophoresis, Western blotting and protein sequencing were used to characterize this protein. For detailed analysis, commercial mouse monoclonal antibody against human sperm-specific glycolytic enzyme (GAPDHS – ab57062, Abcam, UK) was used. Results: MoAb Hs-8 is a human sperm antibody that cross-reacts with the Hs-8-related protein in spermatozoa of other mammalian species (boar, mouse). In the immunofluorescence test Hs-8 antibody reacted with a protein localised in the arcosomal part of the sperm head and in the principal piece of the sperm flagellum. MoAb Hs-8 labelled a protein of 44 kDa in immunoblot test in the extract of human sperm. Sequence analysis characterized protein Hs-8 as GAPDS (glyceraldehyde 3-phosphate dehydrohenase-spermatogenic). For this reason commercial mouse monoclonal antibody against human sperm-specific glycolytic enzyme (GAPDHS) was applied in other tests. Both antibodies labelled similar proteins in immunofluorescence test (the result was proved by colocalization), in electron microscopy and in immunoblot analysis. Conclusion: GAPDHS is a sperm-specific glycolytic enzyme required for energy production during spermatogenesis and for sperm motility. In our previous paper (JP et al, Theriogeology, 2001) we found that monoclonal antibody Hs-8 blocked the sperm-zona pellucida binding. This protein can be important for secondary sperm/oocyte binding. The role of GAPDS in the sperm head has not yet been elucidated.
Trvalý link: http://hdl.handle.net/11104/0217279