Binding of recombinant human proacrosin/acrosin to zona pellucida (ZP) glycoproteins. I. Studies with recombinant human ZPA, ZPB, and ZPC
Objective: To characterize proacrosin/acrosin interaction with isolated zona pellucida (ZP) components. 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 from human ZP glycopro...
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todo:paper_00150282_v83_n6_p1780_Furlong2023-10-03T14:13:27Z Binding of recombinant human proacrosin/acrosin to zona pellucida (ZP) glycoproteins. I. Studies with recombinant human ZPA, ZPB, and ZPC Furlong, L.I. Harris, J.D. Vazquez-Levin, M.H. Acrosin Acrosome Fertilization Human Sperm-egg interaction Spermatozoa Zona pellucida acrosin dextran sulfate fucose glycan derivative glycoprotein mannose proacrosin protein subunit recombinant protein amino terminal sequence article binding affinity binding site carboxy terminal sequence comparative study human nonhuman priority journal protein binding protein function protein interaction zona pellucida Acrosin Animals Binding Sites CHO Cells Cricetinae Egg Proteins Enzyme Precursors Humans Male Membrane Glycoproteins Prospective Studies Protein Binding Receptors, Cell Surface Recombinant Proteins Zona Pellucida Objective: To characterize proacrosin/acrosin interaction with isolated zona pellucida (ZP) components. 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 from human ZP glycoproteins (rec-hZPA, ZPB, and ZPC). Intervention(s): In vitro binding assay developed to assess proacrosin/acrosin-ZP interaction. Main Outcome Measure(s): Zona pellucida glycoprotein binding to proacrosin/acrosin; estimation of binding affinity. Result(s): Of all ZP proteins, rec-hZPA demonstrated the highest binding activity toward acrosin (Rec-30) (rec-hZPB: 42% of rec-hZPA; rec-hZPC: 39% of rec-hZPA; P<.0005). Rec-hZPA interaction was disturbed by dextran sulphate (75% inhibition with 10 μM), fucose (67% inhibition with 1.5 μM), and mannose (69% inhibition with 333 mM). Comparing binding activity of proacrosin with other N-terminal acrosin fragments, Rec-40 showed 2.6-3 times higher levels. Moreover, saturable high affinity binding of Rec-40 to ZP components was observed (Kd: 34 nM for rec-hZPA, 38 nM for rec-hZPB, 63 nM for rec-hZPC). Conclusion(s): The rec-hZPA is the major ZP ligand for human proacrosin/acrosin. The interaction involves mannosyl, fucosyl, and sulfated glycans. Binding sites for rec-hZP would be located both at the N- and C-terminus of proacrosin, revealing a key role of the proenzyme in the interaction. ©2005 by American Society for Reproductive Medicine. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_00150282_v83_n6_p1780_Furlong |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
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R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Acrosin Acrosome Fertilization Human Sperm-egg interaction Spermatozoa Zona pellucida acrosin dextran sulfate fucose glycan derivative glycoprotein mannose proacrosin protein subunit recombinant protein amino terminal sequence article binding affinity binding site carboxy terminal sequence comparative study human nonhuman priority journal protein binding protein function protein interaction zona pellucida Acrosin Animals Binding Sites CHO Cells Cricetinae Egg Proteins Enzyme Precursors Humans Male Membrane Glycoproteins Prospective Studies Protein Binding Receptors, Cell Surface Recombinant Proteins Zona Pellucida |
| spellingShingle |
Acrosin Acrosome Fertilization Human Sperm-egg interaction Spermatozoa Zona pellucida acrosin dextran sulfate fucose glycan derivative glycoprotein mannose proacrosin protein subunit recombinant protein amino terminal sequence article binding affinity binding site carboxy terminal sequence comparative study human nonhuman priority journal protein binding protein function protein interaction zona pellucida Acrosin Animals Binding Sites CHO Cells Cricetinae Egg Proteins Enzyme Precursors Humans Male Membrane Glycoproteins Prospective Studies Protein Binding Receptors, Cell Surface Recombinant Proteins Zona Pellucida Furlong, L.I. Harris, J.D. Vazquez-Levin, M.H. Binding of recombinant human proacrosin/acrosin to zona pellucida (ZP) glycoproteins. I. Studies with recombinant human ZPA, ZPB, and ZPC |
| topic_facet |
Acrosin Acrosome Fertilization Human Sperm-egg interaction Spermatozoa Zona pellucida acrosin dextran sulfate fucose glycan derivative glycoprotein mannose proacrosin protein subunit recombinant protein amino terminal sequence article binding affinity binding site carboxy terminal sequence comparative study human nonhuman priority journal protein binding protein function protein interaction zona pellucida Acrosin Animals Binding Sites CHO Cells Cricetinae Egg Proteins Enzyme Precursors Humans Male Membrane Glycoproteins Prospective Studies Protein Binding Receptors, Cell Surface Recombinant Proteins Zona Pellucida |
| description |
Objective: To characterize proacrosin/acrosin interaction with isolated zona pellucida (ZP) components. 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 from human ZP glycoproteins (rec-hZPA, ZPB, and ZPC). Intervention(s): In vitro binding assay developed to assess proacrosin/acrosin-ZP interaction. Main Outcome Measure(s): Zona pellucida glycoprotein binding to proacrosin/acrosin; estimation of binding affinity. Result(s): Of all ZP proteins, rec-hZPA demonstrated the highest binding activity toward acrosin (Rec-30) (rec-hZPB: 42% of rec-hZPA; rec-hZPC: 39% of rec-hZPA; P<.0005). Rec-hZPA interaction was disturbed by dextran sulphate (75% inhibition with 10 μM), fucose (67% inhibition with 1.5 μM), and mannose (69% inhibition with 333 mM). Comparing binding activity of proacrosin with other N-terminal acrosin fragments, Rec-40 showed 2.6-3 times higher levels. Moreover, saturable high affinity binding of Rec-40 to ZP components was observed (Kd: 34 nM for rec-hZPA, 38 nM for rec-hZPB, 63 nM for rec-hZPC). Conclusion(s): The rec-hZPA is the major ZP ligand for human proacrosin/acrosin. The interaction involves mannosyl, fucosyl, and sulfated glycans. Binding sites for rec-hZP would be located both at the N- and C-terminus of proacrosin, revealing a key role of the proenzyme in the interaction. ©2005 by American Society for Reproductive Medicine. |
| format |
JOUR |
| author |
Furlong, L.I. Harris, J.D. Vazquez-Levin, M.H. |
| author_facet |
Furlong, L.I. Harris, J.D. Vazquez-Levin, M.H. |
| author_sort |
Furlong, L.I. |
| title |
Binding of recombinant human proacrosin/acrosin to zona pellucida (ZP) glycoproteins. I. Studies with recombinant human ZPA, ZPB, and ZPC |
| title_short |
Binding of recombinant human proacrosin/acrosin to zona pellucida (ZP) glycoproteins. I. Studies with recombinant human ZPA, ZPB, and ZPC |
| title_full |
Binding of recombinant human proacrosin/acrosin to zona pellucida (ZP) glycoproteins. I. Studies with recombinant human ZPA, ZPB, and ZPC |
| title_fullStr |
Binding of recombinant human proacrosin/acrosin to zona pellucida (ZP) glycoproteins. I. Studies with recombinant human ZPA, ZPB, and ZPC |
| title_full_unstemmed |
Binding of recombinant human proacrosin/acrosin to zona pellucida (ZP) glycoproteins. I. Studies with recombinant human ZPA, ZPB, and ZPC |
| title_sort |
binding of recombinant human proacrosin/acrosin to zona pellucida (zp) glycoproteins. i. studies with recombinant human zpa, zpb, and zpc |
| url |
http://hdl.handle.net/20.500.12110/paper_00150282_v83_n6_p1780_Furlong |
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