The phospholipase activity of Staphylococcus hyicus lipase strongly depends on a single Ser to Val mutation
| Autor: | Hubertus M. Verheij, M. Van Kampen, Maarten R. Egmond, Niek Dekker, Jan-Willem F. A. Simons |
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| Rok vydání: | 1998 |
| Předmět: |
Stereochemistry
Staphylococcus Molecular Sequence Data Phospholipase medicine.disease_cause Biochemistry Phosphates Substrate Specificity Bacterial Proteins parasitic diseases Serine medicine Point Mutation Amino Acid Sequence Lipase Molecular Biology Phospholipids Staphylococcus hyicus chemistry.chemical_classification Mutation Binding Sites Sequence Homology Amino Acid biology Point mutation Organic Chemistry Mutagenesis Valine Cell Biology biology.organism_classification Amino acid Enzyme chemistry Phospholipases Mutagenesis Site-Directed biology.protein |
| Zdroj: | Chemistry and Physics of Lipids. 93:39-45 |
| ISSN: | 0009-3084 |
| DOI: | 10.1016/s0009-3084(98)00027-9 |
| Popis: | Site-directed mutagenesis and domain exchange were used to investigate the role of the C-terminal domains of Staphylococcus hyicus lipase (SHL) and S. aureus lipase (SAL) in substrate selectivity. The introduction of a single point mutation coding for the substitution of Val for Ser356 in SHL yields an enzyme which has retained full lipase activity, although with more than 12-fold lower phospholipase activity. Starting with this S356V variant of SHL the C-terminal 40 amino acids were replaced by the corresponding SAL sequence. Although 23 amino acid changes were introduced simultaneously the impact on the phospholipase/lipase activity ratio was only 4-fold. We therefore conclude that in the C-terminal domain it is Ser356 which mainly determines phospholipase activity. The introduction of a Val357 to Ser substitution in SAL did not turn SAL into a phospholipase, showing that residues from other domains contribute to this activity as well. The results are discussed in view of the sequence homology of lipases and (lyso)phospholipases. |
| Databáze: | OpenAIRE |
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