| Thimet oligopeptidase: site-directed mutagenesis disproves previous assumptions about the nature of the catalytic site. | |
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MedLine Citation:
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PMID: 9755850 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Zinc metallopeptidases that contain the His-Glu-Xaa-Xaa-His (HEXXH) motif generally have a third ligand of the metal ion that may be either a Glu residue (in clan MA) or a His residue (in clan MB) (Rawlings and Barrett (1995) Methods Enzymol. 248, 183-228). Thimet oligopeptidase has not yet been assigned to either clan, and both Glu and His residues have been proposed as the third ligand. We mutated candidate ligand residues in the recombinant enzyme and identified Glu, His and Asp residues that are important for catalytic activity and/or stability of the protein. However, neither of the Glu and His residues close to the HEXXH motif that have previously been suggested to be ligands is required for the binding of zinc. We conclude that thimet oligopeptidase is not a member of clan MA or clan MB and it is likely that the enzyme possesses a catalytic site and protein fold different from those identified in any metallopeptidase to date. The definitive identification of the third zinc ligand may well require the determination of the crystallographic structure of thimet oligopeptidase or one of its homologues. |
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Authors:
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J M Chen; R A Stevens; P W Wray; N D Rawlings; A J Barrett |
Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: FEBS letters Volume: 435 ISSN: 0014-5793 ISO Abbreviation: FEBS Lett. Publication Date: 1998 Sep |
Date Detail:
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Created Date: 1998-10-05 Completed Date: 1998-10-05 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 0155157 Medline TA: FEBS Lett Country: NETHERLANDS |
Other Details:
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Languages: eng Pagination: 16-20 Citation Subset: IM |
Affiliation:
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MRC Peptidase Laboratory, The Babraham Institute, Cambridge, UK. Jinq-May.Chen@bbsrc.ac.uk |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Amino Acid Sequence Animals Binding Sites / genetics Catalysis Enzyme Activation / genetics Gene Expression Regulation, Enzymologic Glutamic Acid / genetics Histidine / genetics Ligands Metalloendopeptidases / biosynthesis, genetics*, metabolism Molecular Sequence Data Mutagenesis, Site-Directed* Rats Recombinant Proteins / biosynthesis, metabolism Zinc / metabolism |
| Chemical | |
Reg. No./Substance:
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0/Ligands; 0/Recombinant Proteins; 56-86-0/Glutamic Acid; 71-00-1/Histidine; 7440-66-6/Zinc; EC 3.4.24.-/Metalloendopeptidases; EC 3.4.24.15/thimet oligopeptidase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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