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dc.contributor.author
Ellerman, Diego Andrés
dc.contributor.author
Da Ros, Vanina Gabriela
dc.contributor.author
Cohen, Debora Juana
dc.contributor.author
Busso, Dolores
dc.contributor.author
Morgenfeld, Mauro Miguel
dc.contributor.author
Cuasnicu, Patricia Sara
dc.date.available
2017-12-22T05:25:28Z
dc.date.issued
2002-04
dc.identifier.citation
Ellerman, Diego Andrés; Da Ros, Vanina Gabriela; Cohen, Debora Juana; Busso, Dolores; Morgenfeld, Mauro Miguel; et al.; Expression and structure-function analysis of de, a sperm cysteine rich secretory protein that mediates gamete fusion; Society for the Study of Reproduction; Biology of Reproduction; 67; 4; 4-2002; 1225-1231
dc.identifier.issn
0006-3363
dc.identifier.uri
http://hdl.handle.net/11336/31339
dc.description.abstract
Rat sperm epididymal glycoprotein DE belongs to the cysteine- rich secretory protein (CRISP) family and participates in sperm-egg fusion through its binding to complementary sites on the egg surface. To investigate the molecular mechanisms underlying the role of DE in gamete fusion, in the present work we expressed DE in a prokaryotic system, and examined the relevance of carbohydrates and disulfide bonds for the biological activity of the protein. Immunofluorescence and sperm-egg fusion assays carried out in the presence of recombinant DE (recDE) revealed that this protein exhibits the ability to bind to the DE-egg binding sites and to inhibit gamete fusion, as does native DE (nDE). Comparison of the proteins indicated, however, that the inhibitory ability of recDE was significantly lower than that of nDE. This difference would not be due to the lack of carbohydrates in the bacterially expressed protein because enzymatically deglycosylated nDE was as able as the untreated protein to inhibit gamete fusion. To examine whether disulfide bridges are involved in DE activity, the presence of sulfhydryls in nDE and recDE was evaluated by the biotin-maleimide technique. Results indicated that, unlike nDE, in which all cysteines are involved in disulfide bonds, recDE contains free thiol groups. Subsequent experiments showed that reduction of nDE with dithiothreitol significantly decreased the ability of the protein to inhibit gamete fusion. Together, these results indicate that whereas carbohydrates do not have a role in DE-mediated gamete fusion, disulfide bridges are required for full biological activity of the protein. To our knowledge, this is the first study reporting the relevance of structural components for the function of a CRISP member.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Society for the Study of Reproduction
dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.subject
Epididymis
dc.subject
Fer4tilization
dc.subject
Fusion
dc.subject
Ovum
dc.subject
Sperm
dc.subject
Epididymis
dc.subject.classification
Biología Reproductiva
dc.subject.classification
Ciencias Biológicas
dc.subject.classification
CIENCIAS NATURALES Y EXACTAS
dc.title
Expression and structure-function analysis of de, a sperm cysteine rich secretory protein that mediates gamete fusion
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2017-12-04T17:55:27Z
dc.identifier.eissn
1529-7268
dc.journal.volume
67
dc.journal.number
4
dc.journal.pagination
1225-1231
dc.journal.pais
Estados Unidos
dc.description.fil
Fil: Ellerman, Diego Andrés. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
dc.description.fil
Fil: Da Ros, Vanina Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
dc.description.fil
Fil: Cohen, Debora Juana. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
dc.description.fil
Fil: Busso, Dolores. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
dc.description.fil
Fil: Morgenfeld, Mauro Miguel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
dc.description.fil
Fil: Cuasnicu, Patricia Sara. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
dc.journal.title
Biology of Reproduction
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/https://academic.oup.com/biolreprod/article/67/4/1225/2683553
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1095/biolreprod67.4.1225
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