Site-directed mutagenesis of boar proacrosin reveals residues involved in binding of zona pellucida glycoproteins
| Autor: | Bertram Brenig, Ingo Jenneckens, John Coadwell, B. Kriegesmann, Brit Marschall, Stephan Jansen, Roy Jones |
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| Rok vydání: | 1998 |
| Předmět: |
Male
Models Molecular Zona pellucida glycoprotein Protein Conformation Swine Recombinant Fusion Proteins Receptors Cell Surface Biology Zona Pellucida Glycoproteins Genetics medicine Animals Binding site Zona pellucida Site-directed mutagenesis chemistry.chemical_classification Serine protease Acrosin Enzyme Precursors Binding Sites Membrane Glycoproteins Egg Proteins Cell Biology Amino acid medicine.anatomical_structure Biochemistry chemistry biology.protein Mutagenesis Site-Directed Glycoprotein Developmental Biology |
| Zdroj: | Molecular reproduction and development. 51(2) |
| ISSN: | 1040-452X |
| Popis: | Proacrosin, the zymogen form of the serine protease β-acrosin, is thought to function as a secondary binding molecule between mammalian gametes during fertilization (Jansen et al., 1995: Int J Dev Biol 39, 501–510). The interaction involves strong ionic bonds between positively charged amino acids on proacrosin and negatively charged polysulphate groups on zona pellucida glycoproteins. In this investigation, we identified the basic residues on proacrosin that are important for this binding. Site-directed mutagenesis shows that two groups of amino acids comprising His47, Arg50, and Arg51 together with Arg250, Lys252, and Arg253 are crucial because their deletion or replacement severely reduces affinity for zona glycoproteins. Molecular models of proacrosin reveal that these residues are located along one face of the protein on two exposed surface loops that project over and around the catalytic site. These findings support the hypothesis that polysulphate binding sites on proacrosin are formed by a restricted number of basic amino acids on the surface of the protein, presenting a specific orientation that is complementary to negatively charged sulphate groups on zona glycoproteins. Identification and elucidation of the stereochemistry of these charged moieties will aid design of new kinds of nonsteroidal antifertility agents. Mol. Reprod. Dev. 51:184–192, 1998. © 1998 Wiley-Liss, Inc. |
| Databáze: | OpenAIRE |
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