| Genetic and biochemical properties of an extracellular neutral metalloprotease from Staphylococcus hyicus subsp. hyicus. | |
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MedLine Citation:
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PMID: 8121397 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The gene encoding the extracellular neutral metalloprotease ShpI from Staphylococcus hyicus subsp. hyicus was cloned. DNA sequencing revealed an ORF of 1317 nucleotides encoding a 438 amino acid protein with Mr of 49,698. When the cloned gene was expressed in Staphylococcus carnosus, a 42 kDa protease was found in the culture medium. The protease was purified from both S. carnosus (pCAshp1) and S. hyicus subsp. hyicus. The N-terminal amino acid sequences of the two proteases revealed that ShpI is organized as a pre-pro-enzyme with a proposed 26 amino acid signal peptide, a 75 amino acid hydrophilic pro-region, and a 337 amino acid extracellular mature form with a calculated Mr of 38,394. The N-termini showed microheterogeneity in both host strains. ShpI had a maximum proteolytic activity at 55 degrees C and pH 7.4-8.5. The protease, which had a low substrate specificity, could be inhibited by metal- and zinc-specific inhibitors, such as EDTA and 1,10-phenanthroline. Insensitivity to phosphoramidon separates ShpI from the thermolysin-like family. The conserved Zn2+ binding motif, the only homology to other proteases, and the reactivation of the apoenzyme by Zn2+, indicated that Zn2+ is the catalytic ion. Ca2+ very probably acts as a stabilizer. We also demonstrated the presence of a second extracellular protease in S. hyicus subsp. hyicus. |
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Authors:
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S Ayora; F Götz |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Molecular & general genetics : MGG Volume: 242 ISSN: 0026-8925 ISO Abbreviation: Mol. Gen. Genet. Publication Date: 1994 Feb |
Date Detail:
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Created Date: 1994-04-06 Completed Date: 1994-04-06 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 0125036 Medline TA: Mol Gen Genet Country: GERMANY |
Other Details:
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Languages: eng Pagination: 421-30 Citation Subset: IM |
Affiliation:
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Universität Tübingen, Germany. |
| Data Bank Information | |
Bank Name/Acc. No.:
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GENBANK/X73315 |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Amino Acid Sequence Base Sequence Cloning, Molecular Genes, Bacterial Hydrogen-Ion Concentration Metalloendopeptidases / genetics*, isolation & purification, metabolism Molecular Sequence Data Restriction Mapping Sequence Homology, Nucleic Acid Staphylococcus / enzymology*, genetics Substrate Specificity Temperature |
| Chemical | |
Reg. No./Substance:
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EC 3.4.24.-/Metalloendopeptidases; EC 3.4.24.-/ShpI metalloprotease |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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