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Abstract:

Objective: To assess the interaction of human proacrosin/acrosin with mannose residues coupled to a protein backbone. Design: Prospective study. Setting: Basic research laboratory. Patient(s): Recombinant proteins derived from human proacrosin (Rec-40, Rec-30, Rec-20, Rec-10, and Rec-6) and bovine serum albumin (BSA)-mannose as ligand. Intervention(s): In vitro binding assay developed to assess proacrosin/acrosin-BSA-mannose interaction. Main Outcome Measure(s): Proacrosin/acrosin binding to BSA-mannose; estimation of binding affinity. Result(s): All recombinant proteins of acrosin but Rec-6 (residues 1-59 of proacrosin) specifically bound to BSA-mannose. Rec-40 (proacrosin) showed the highest binding affinity (dissociation constant Kd, 162 nM), followed by N-terminal fragments Rec-30 (248 nM), Rec-20 (359 nM), and Rec-10 (402 nM). A significant decrease in binding activity was observed when acrosin proteins were treated with denaturing agents such as urea or heat. The β-mercaptoethanol treatment produced a 39% decrease on Rec-30 binding to BSA-mannose; in contrast, no effect was observed with Rec-40, suggesting the presence of at least two types of mannose-binding sites. Conclusion(s): [1] Proacrosin interacts with mannose residues through binding sites located at both the N- and C-terminal portion of the protein, [2] the full-length protein is required for maximal BSA-mannose binding, and [3] binding sites are stabilized by noncovalent bonds and by disulfide linkages. ©2005 by American Society for Reproductive Medicine.

Registro:

Documento: Artículo
Título:Binding of recombinant human proacrosin/acrosin to zona pellucida glycoproteins. II. Participation of mannose residues in the interaction
Autor:Furlong, L.I.; Veaute, C.; Vazquez-Levin, M.H.
Filiación:Instituto de Biología Y Medicina Experimental-CONICET-UBA, Buenos Aires, Argentina
Instituto de Biología Y Medicina Experimental, Vuelta de Obligado 2490, 1428, Buenos Aires, Argentina
Palabras clave:Acrosin; Acrosome; Fertilization; Human; Mannose; Sperm-egg interaction; Spermatozoa; Zona pellucida; acrosin; bovine serum albumin; glycoprotein; mannose; mercaptoethanol; proacrosin; recombinant protein; urea; amino terminal sequence; article; binding affinity; binding site; carboxy terminal sequence; chemical bond; heat; priority journal; prospective study; protein binding; protein interaction; zona pellucida; Acrosin; Animals; Cattle; Egg Proteins; Enzyme Precursors; Humans; Male; Mannose; Membrane Glycoproteins; Prospective Studies; Protein Binding; Protein Interaction Mapping; Receptors, Cell Surface; Recombinant Proteins
Año:2005
Volumen:83
Número:6
Página de inicio:1791
Página de fin:1796
DOI: http://dx.doi.org/10.1016/j.fertnstert.2004.12.043
Título revista:Fertility and Sterility
Título revista abreviado:Fertil. Steril.
ISSN:00150282
CODEN:FESTA
CAS:acrosin, 9068-57-9; mannose, 31103-86-3, 3458-28-4; mercaptoethanol, 60-24-2; proacrosin, 56379-46-5; urea, 57-13-6; Acrosin, EC 3.4.21.10; Egg Proteins; Enzyme Precursors; Mannose, 31103-86-3; Membrane Glycoproteins; proacrosin, EC 3.4.21.-; Receptors, Cell Surface; Recombinant Proteins; zona pellucida glycoproteins; ZP4 protein, human
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00150282_v83_n6_p1791_Furlong

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Citas:

---------- APA ----------
Furlong, L.I., Veaute, C. & Vazquez-Levin, M.H. (2005) . Binding of recombinant human proacrosin/acrosin to zona pellucida glycoproteins. II. Participation of mannose residues in the interaction. Fertility and Sterility, 83(6), 1791-1796.
http://dx.doi.org/10.1016/j.fertnstert.2004.12.043
---------- CHICAGO ----------
Furlong, L.I., Veaute, C., Vazquez-Levin, M.H. "Binding of recombinant human proacrosin/acrosin to zona pellucida glycoproteins. II. Participation of mannose residues in the interaction" . Fertility and Sterility 83, no. 6 (2005) : 1791-1796.
http://dx.doi.org/10.1016/j.fertnstert.2004.12.043
---------- MLA ----------
Furlong, L.I., Veaute, C., Vazquez-Levin, M.H. "Binding of recombinant human proacrosin/acrosin to zona pellucida glycoproteins. II. Participation of mannose residues in the interaction" . Fertility and Sterility, vol. 83, no. 6, 2005, pp. 1791-1796.
http://dx.doi.org/10.1016/j.fertnstert.2004.12.043
---------- VANCOUVER ----------
Furlong, L.I., Veaute, C., Vazquez-Levin, M.H. Binding of recombinant human proacrosin/acrosin to zona pellucida glycoproteins. II. Participation of mannose residues in the interaction. Fertil. Steril. 2005;83(6):1791-1796.
http://dx.doi.org/10.1016/j.fertnstert.2004.12.043