Document Detail


Genetic and biochemical properties of an extracellular neutral metalloprotease from Staphylococcus hyicus subsp. hyicus.
MedLine Citation:
PMID:  8121397     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
S Ayora; F Götz
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Molecular & general genetics : MGG     Volume:  242     ISSN:  0026-8925     ISO Abbreviation:  Mol. Gen. Genet.     Publication Date:  1994 Feb 
Date Detail:
Created Date:  1994-04-06     Completed Date:  1994-04-06     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0125036     Medline TA:  Mol Gen Genet     Country:  GERMANY    
Other Details:
Languages:  eng     Pagination:  421-30     Citation Subset:  IM    
Affiliation:
Universität Tübingen, Germany.
Data Bank Information
Bank Name/Acc. No.:
GENBANK/X73315
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MeSH Terms
Descriptor/Qualifier:
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:
EC 3.4.24.-/Metalloendopeptidases; EC 3.4.24.-/ShpI metalloprotease

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