The importance of glycine-30 for enzymatic activity of phospholipase A2
| Autor: | Gerard H. de Haas, Peet A. Franken, Hubertus M. Verheij, Ellis Toxopeus, August C.A.P.A. Bekkers |
|---|---|
| Rok vydání: | 1991 |
| Předmět: |
Swine
Molecular Sequence Data Restriction Mapping Mutant Glycine Biophysics Biochemistry Phospholipases A Cofactor Substrate Specificity Serine Phospholipase A2 Structural Biology Animals Calcium ion binding Pancreas Molecular Biology chemistry.chemical_classification Phospholipase A Binding Sites Base Sequence biology Chemistry Kinetics Phospholipases A2 Enzyme Snake venom Mutagenesis Site-Directed biology.protein Oligonucleotide Probes |
| Zdroj: | Biochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology. 1076:374-378 |
| ISSN: | 0167-4838 |
| DOI: | 10.1016/0167-4838(91)90479-j |
| Popis: | The nearly conserved glycine-30 in porcine pancreatic phospholipase A2 has been replaced by serine. The resulting mutant G30S was expressed in Escherichia coli, purified and characterized. The mutation caused a significant drop in enzymatic activity towards monomeric and aggregated substrates, but had a limited effect on substrate binding. In contrast the affinity for calcium ions, the essential cofactor, was reduced 10-fold. The reduced enzymatic activity is attributed to a reduced stabilization of the transition state. The results are discussed in view of naturally occurring inactive phospholipase A2 homologues from snake venom. |
| Databáze: | OpenAIRE |
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