Cloning and characterization of a cysteine proteinase from Saccharomyces cerevisiae
| Autor: | N G, Kambouris, D J, Burke, C E, Creutz |
|---|---|
| Rok vydání: | 1992 |
| Předmět: |
Saccharomyces cerevisiae Proteins
Base Sequence Glycoside Hydrolases Sequence Homology Amino Acid Genes Fungal Molecular Sequence Data Restriction Mapping Saccharomyces cerevisiae Aminopeptidases Substrate Specificity DNA-Binding Proteins Cysteine Endopeptidases Cysteine Proteases Amino Acid Sequence Cloning Molecular DNA Fungal Sequence Alignment |
| Zdroj: | The Journal of biological chemistry. 267(30) |
| ISSN: | 0021-9258 |
| Popis: | We have isolated a gene from Saccharomyces cerevisiae that encodes a protein homologous to the mammalian cysteine proteinase bleomycin hydrolase. Sequence comparison between the yeast and rabbit proteins indicates an amino acid identity of 41.5% over 277 residues and a similarity of 78.3% when conservative substitutions are included. The apparent mass of the yeast protein by sodium dodecyl sulfate-polyacrylamide gel electrophoresis is 47 kDa, although sequence analysis indicates two potential initiator methionines that suggest calculated masses of either 51 or 55 kDa. The protein is nonessential in yeast as haploid mutants disrupted at several positions along the open reading frame remain viable. Furthermore, these mutants do not exhibit any readily observable growth defects under varying conditions of temperature, nutrients, osmotic strength, or exogenous bleomycin. However, the purified protein does exhibit marked hydrolytic activity toward the substrate arginine 4-methyl-7-coumarylamide (Km = 12.8 microM, Vmax = 2.56 mumol mg-1 h-1), and yeast cells engineered to express this protein at higher levels maintain increased resistance to bleomycin compared to wild-type cells. Because this protein represents the first example of a cysteine proteinase identified in yeast, we have named it Ycp1 (yeast cysteine proteinase). |
| Databáze: | OpenAIRE |
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