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dc.contributor.author
Furlong, Laura Ines
dc.contributor.author
Hellman, Ulf
dc.contributor.author
Krimer, Alejandro
dc.contributor.author
Tezon, Jorge Guillermo
dc.contributor.author
Charreau, Eduardo Hernan
dc.contributor.author
Vazquez, Monica Hebe
dc.date.available
2017-12-21T13:48:40Z
dc.date.issued
2000
dc.identifier.citation
Furlong, Laura Ines; Hellman, Ulf; Krimer, Alejandro; Tezon, Jorge Guillermo; Charreau, Eduardo Hernan; et al.; Expression of human proacrosin in Escherichia coli and binding to zona pellucida; Society for the Study of Reproduction; Biology of Reproduction; 62; 3; 2000; 606-615
dc.identifier.issn
0006-3363
dc.identifier.uri
http://hdl.handle.net/11336/31193
dc.description.abstract
Proacrosin is a multifunctional protein present in the sperm acrosome. This study characterizes the expression of human proacrosin in bacteria and assesses zona pellucida binding activity. The cDNA encoding human proacrosin was subcloned in pGEX-3X and pET-22b vectors. In the pGEX system, expression of the full-length fusion protein was not detected. In the pET system, an expression product with an apparent molecular size similar to that expected for the proenzyme (Rec-40, 42-44 kDa) was recognized by a monoclonal antibody to human acrosin, AcrC5F10. A 32-34-kDa protein (Rec-30), not recognized by AcrC5F10 on Western blots, was the major expression product. Proteins of 21 (Rec-20) and 18 (Rec-10) kDa were recovered as insoluble expression products as were Rec-40 and Rec-30, and truncated products from the C terminus were detected in the soluble fraction. Rec-40 and Rec-30 coexisted at any culture time tested. Immune serum raised against Rec-30 (AntiRec-30) stained the acrosomal region of permeabilized human spermatozoa and recognized the recombinant proteins and proacrosin from human sperm extracts. Amino acid sequence analysis indicated that Rec-30, Rec-20, and Rec-10 are N-terminal fragments of proacrosin. The recombinant proteins Rec-40, -30, -20, and -10 were found to interact with homologous (125)I-zona pellucida glycoproteins.
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
Acrosin
dc.subject
Enzyme Precursors
dc.subject
Escherichia Coli
dc.subject
Genetic Vectors
dc.subject
Zona Pellucida
dc.subject.classification
Otras Ciencias Biológicas
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Ciencias Biológicas
dc.subject.classification
CIENCIAS NATURALES Y EXACTAS
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Patología
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Medicina Básica
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CIENCIAS MÉDICAS Y DE LA SALUD
dc.title
Expression of human proacrosin in Escherichia coli and binding to zona pellucida
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2017-12-04T17:54:16Z
dc.identifier.eissn
1529-7268
dc.journal.volume
62
dc.journal.number
3
dc.journal.pagination
606-615
dc.journal.pais
Estados Unidos
dc.journal.ciudad
Nueva York
dc.description.fil
Fil: Furlong, Laura Ines. 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: Hellman, Ulf. Uppasala Branch Biomedical Center; Suiza
dc.description.fil
Fil: Krimer, Alejandro. Biosidus; Argentina
dc.description.fil
Fil: Tezon, Jorge Guillermo. 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: Charreau, Eduardo Hernan. 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: Vazquez, Monica Hebe. 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/62/3/606/2734731
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1095/biolreprod62.3.606
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