Site-directed mutagenesis of a calcium binding site modifies specifically the different biochemical properties of annexin I
| Autor: | Joannes Sri Widada, G. Travé, David Cregut, J.-F. Quignard, Corinne Lionne, Jean-Pierre Liautard, Laurent Chiche |
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| Přispěvatelé: | Centre National de la Recherche Scientifique (CNRS) |
| Rok vydání: | 1994 |
| Předmět: |
Models
Molecular Protein Folding [SDV]Life Sciences [q-bio] Mutant Glycine Glutamic Acid chemistry.chemical_element Mutagenesis (molecular biology technique) Bioengineering Calcium Biology Biochemistry law.invention Structure-Activity Relationship 03 medical and health sciences Drug Stability Glutamates law Escherichia coli Consensus sequence Binding site Site-directed mutagenesis Molecular Biology ComputingMilieux_MISCELLANEOUS Annexin A1 030304 developmental biology 0303 health sciences Binding Sites Molecular Structure 030302 biochemistry & molecular biology Recombinant Proteins chemistry Liposomes Mutagenesis Site-Directed Recombinant DNA Thermodynamics Dialysis Biotechnology Binding domain |
| Zdroj: | Protein Engineering, Design and Selection Protein Engineering, Design and Selection, Oxford University Press (OUP), 1994, 7 (5), pp.689-696. ⟨10.1093/protein/7.5.689⟩ |
| ISSN: | 1741-0134 1741-0126 |
| DOI: | 10.1093/protein/7.5.689 |
| Popis: | All the functions of annexins in vitro as well as in vivo are mediated and probably regulated by calcium. We have used recombinant annexin I, synthesized by Escherichia coli, and we have performed site-directed mutagenesis. We have mutated the endonexin fold of domain 2 that binds calcium. Mutations were performed in this domain of the molecule because it perfectly matches the calcium binding consensus sequence. The two glycines of this fold were mutated into glutamic acid. The helix content and the stability of the mutants are identical to those of the wild-type, suggesting that the mutations did not drastically affect the structure of the protein. The two mutants showed modified calcium binding affinities. However, the calcium binding affinity of the G131E mutant was far more altered than that of the G129E mutant. Furthermore, other biochemical properties of these mutants were modified to different extents. The binding to phospholipid was not seriously affected, whereas the self-association was lost by the G131E mutant. In the same way, liposome aggregation is conserved, but modified, while the calcium affinity measured by equilibrium dialysis is dramatically altered. |
| Databáze: | OpenAIRE |
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