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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Detalles Bibliográficos
Autores principales: Furlong, L.I., Harris, J.D., Vazquez-Levin, M.H.
Formato: JOUR
Materias:
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
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
Acceso en línea:http://hdl.handle.net/20.500.12110/paper_00150282_v83_n6_p1780_Furlong
Aporte de:
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires
Descripción
Sumario: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.

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