Binding of recombinant human proacrosin/acrosin to zona pellucida glycoproteins. II. Participation of mannose residues in the interaction
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 s...
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2005
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00150282_v83_n6_p1791_Furlong http://hdl.handle.net/20.500.12110/paper_00150282_v83_n6_p1791_Furlong |
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paper:paper_00150282_v83_n6_p1791_Furlong2023-06-08T14:37:56Z Binding of recombinant human proacrosin/acrosin to zona pellucida glycoproteins. II. Participation of mannose residues in the interaction 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 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. 2005 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00150282_v83_n6_p1791_Furlong http://hdl.handle.net/20.500.12110/paper_00150282_v83_n6_p1791_Furlong |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
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 |
| spellingShingle |
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 Binding of recombinant human proacrosin/acrosin to zona pellucida glycoproteins. II. Participation of mannose residues in the interaction |
| topic_facet |
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 |
| description |
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. |
| title |
Binding of recombinant human proacrosin/acrosin to zona pellucida glycoproteins. II. Participation of mannose residues in the interaction |
| title_short |
Binding of recombinant human proacrosin/acrosin to zona pellucida glycoproteins. II. Participation of mannose residues in the interaction |
| title_full |
Binding of recombinant human proacrosin/acrosin to zona pellucida glycoproteins. II. Participation of mannose residues in the interaction |
| title_fullStr |
Binding of recombinant human proacrosin/acrosin to zona pellucida glycoproteins. II. Participation of mannose residues in the interaction |
| title_full_unstemmed |
Binding of recombinant human proacrosin/acrosin to zona pellucida glycoproteins. II. Participation of mannose residues in the interaction |
| title_sort |
binding of recombinant human proacrosin/acrosin to zona pellucida glycoproteins. ii. participation of mannose residues in the interaction |
| publishDate |
2005 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00150282_v83_n6_p1791_Furlong http://hdl.handle.net/20.500.12110/paper_00150282_v83_n6_p1791_Furlong |
| _version_ |
1768546330414678016 |