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Statements

Subject Item
n2:RIV%2F86652036%3A_____%2F12%3A00380394%21RIV13-AV0-86652036
rdf:type
skos:Concept n14:Vysledek
dcterms:description
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. 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.
dcterms:title
Characterization of human sperm protein recognized by monoclonal antibody HS-8 Characterization of human sperm protein recognized by monoclonal antibody HS-8
skos:prefLabel
Characterization of human sperm protein recognized by monoclonal antibody HS-8 Characterization of human sperm protein recognized by monoclonal antibody HS-8
skos:notation
RIV/86652036:_____/12:00380394!RIV13-AV0-86652036
n14:predkladatel
n15:ico%3A86652036
n3:aktivita
n17:Z n17:P
n3:aktivity
P(GA523/09/1793), P(GAP503/12/1834), Z(AV0Z50520701)
n3:dodaniDat
n4:2013
n3:domaciTvurceVysledku
n16:9747060 n16:4192567 n16:8739927
n3:druhVysledku
n18:O
n3:duvernostUdaju
n13:S
n3:entitaPredkladatele
n11:predkladatel
n3:idSjednocenehoVysledku
126817
n3:idVysledku
RIV/86652036:_____/12:00380394
n3:jazykVysledku
n8:eng
n3:klicovaSlova
monoclonal antibody; GAPDHS; human sperm proteins; mass spectrometry
n3:klicoveSlovo
n5:human%20sperm%20proteins n5:mass%20spectrometry n5:GAPDHS n5:monoclonal%20antibody
n3:kontrolniKodProRIV
[33D4AA1406C9]
n3:obor
n10:EC
n3:pocetDomacichTvurcuVysledku
3
n3:pocetTvurcuVysledku
3
n3:projekt
n12:GA523%2F09%2F1793 n12:GAP503%2F12%2F1834
n3:rokUplatneniVysledku
n4:2012
n3:tvurceVysledku
Pěknicová, Jana Margaryan, Hasmik Čapková, Jana
n3:zamer
n9:AV0Z50520701