Artículo

Furlong, L.I.; Hellman, U.; Krimer, A.; Tezón, J.G.; Charreau, E.H.; Vazquez-Levin, M.H. "Expression of human proacrosin in Escherichia coli and binding to zona pellucida" (2000) Biology of Reproduction. 62(3):606-615
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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 125I-zona pellucida glycoproteins.

Registro:

Documento: Artículo
Título:Expression of human proacrosin in Escherichia coli and binding to zona pellucida
Autor:Furlong, L.I.; Hellman, U.; Krimer, A.; Tezón, J.G.; Charreau, E.H.; Vazquez-Levin, M.H.
Filiación:Inst. de Biología y Medicina, Exp.-Consejo Nac. Invest. Cie. T., (1428) Buenos Aires, Argentina
Ludwig Institute for Cancer Research, Uppsala Branch Biomedical Center, S-751 24 Uppsala, Sweden
Biosidus, Buenos Aires, Argentina
Inst. de Biología y Medicina, Exp.-Consejo Nac. Invest. Cie. T., Vuelta de Obligado 2490, (1428) Buenos Aires, Argentina
Palabras clave:complementary DNA; epitope; glycoprotein; iodine 125; polyclonal antibody; proacrosin; recombinant protein; article; controlled study; DNA sequence; Escherichia coli; gene expression; human; human cell; male; nonhuman; polyacrylamide gel electrophoresis; polymerase chain reaction; priority journal; protein expression; zona pellucida; Escherichia coli
Año:2000
Volumen:62
Número:3
Página de inicio:606
Página de fin:615
DOI: http://dx.doi.org/10.1095/biolreprod62.3.606
Título revista:Biology of Reproduction
Título revista abreviado:Biol. Reprod.
ISSN:00063363
CODEN:BIREB
CAS:iodine 125, 14158-31-7, 22822-81-7; proacrosin, 56379-46-5
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00063363_v62_n3_p606_Furlong

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Citas:

---------- APA ----------
Furlong, L.I., Hellman, U., Krimer, A., Tezón, J.G., Charreau, E.H. & Vazquez-Levin, M.H. (2000) . Expression of human proacrosin in Escherichia coli and binding to zona pellucida. Biology of Reproduction, 62(3), 606-615.
http://dx.doi.org/10.1095/biolreprod62.3.606
---------- CHICAGO ----------
Furlong, L.I., Hellman, U., Krimer, A., Tezón, J.G., Charreau, E.H., Vazquez-Levin, M.H. "Expression of human proacrosin in Escherichia coli and binding to zona pellucida" . Biology of Reproduction 62, no. 3 (2000) : 606-615.
http://dx.doi.org/10.1095/biolreprod62.3.606
---------- MLA ----------
Furlong, L.I., Hellman, U., Krimer, A., Tezón, J.G., Charreau, E.H., Vazquez-Levin, M.H. "Expression of human proacrosin in Escherichia coli and binding to zona pellucida" . Biology of Reproduction, vol. 62, no. 3, 2000, pp. 606-615.
http://dx.doi.org/10.1095/biolreprod62.3.606
---------- VANCOUVER ----------
Furlong, L.I., Hellman, U., Krimer, A., Tezón, J.G., Charreau, E.H., Vazquez-Levin, M.H. Expression of human proacrosin in Escherichia coli and binding to zona pellucida. Biol. Reprod. 2000;62(3):606-615.
http://dx.doi.org/10.1095/biolreprod62.3.606