| Activation of an Asp-124-->Asn mutant of haloalkane dehalogenase by hydrolytic deamidation of asparagine. | |
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MedLine Citation:
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PMID: 7828730 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Haloalkane dehalogenase hydrolyses various 1-halo-n-alkanes to the corresponding alcohols by covalent catalysis with formation of an alkyl-enzyme intermediate. The carboxylate function of the nucleophilic aspartate (Asp-124) that displaces the halogen during formation of the intermediate was changed to an amide by site-directed mutagenesis (Asp-124-->Asn). Activity measurements and analysis of peptides containing the nucleophilic residue showed that the mutant enzyme was inactive, but that the activity increased by rapid deamidation of the asparagine residue, yielding wild type enzyme. There was no indication for isoaspartate formation during this process. The results suggest that a water molecule that is located close to the carboxyl function of Asp-124 in the X-ray structure is highly reactive and is responsible for the observed deamidation. |
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Authors:
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F Pries; J Kingma; D B Janssen |
Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: FEBS letters Volume: 358 ISSN: 0014-5793 ISO Abbreviation: FEBS Lett. Publication Date: 1995 Jan |
Date Detail:
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Created Date: 1995-02-23 Completed Date: 1995-02-23 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: 171-4 Citation Subset: IM |
Affiliation:
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Department of Biochemistry, Groningen Biomolecular and Biotechnology Institute, University of Groningen, The Netherlands. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Amides
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chemistry Amino Acid Sequence Asparagine / metabolism* Aspartic Acid / metabolism* Base Sequence DNA Primers Deamination Enzyme Activation Hydrolases / genetics, metabolism* Hydrolysis Molecular Sequence Data Mutation Point Mutation |
| Chemical | |
Reg. No./Substance:
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0/Amides; 0/DNA Primers; 56-84-8/Aspartic Acid; 7006-34-0/Asparagine; EC 3.-/Hydrolases; EC 3.8.1.5/haloalkane dehalogenase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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