| 2-Oxo-3-alkynoic acids, universal mechanism-based inactivators of thiamin diphosphate-dependent decarboxylases: synthesis and evidence for potent inactivation of the pyruvate dehydrogenase multienzyme complex. | |
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MedLine Citation:
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PMID: 9201955 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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A new class of compounds, the 2-oxo-3-alkynoic acids with a phenyl substituent at carbon 4 was reported by the authors as potent irreversible and mechanism-based inhibitors of the thiamin diphosphate- (ThDP-) dependent enzyme pyruvate decarboxylase [Chiu, C.-F., & Jordan, F. (1994) J. Org. Chem. 59, 5763-5766]. The method has been successfully extended to the synthesis of the 4-, 5-, and 7-carbon aliphatic members of this family of compounds. These three compounds were then tested on three ThDP-dependent pyruvate decarboxylases: the Escherichia coli pyruvate dehydrogenase multienzyme complex (PDHc) and its E1 (ThDP-dependent) component, pyruvate oxidase (POX, phosphorylating; from Lactobacillus plantarum),and pyruvate decarboxylase (PDC) from Saccharomycescerevisiae. All three enzymes were irreversibly inhibited by the new compounds. The 4-carbon acid is the best substrate-analog inactivator known to date for PDHc, more potent than either fluoropyruvate or bromopyruvate. The following conclusions were drawn from extensive studies with PDHc: (a) The kinetics of inactivation of PDH complexes and of resolved E1 by 2-oxo-3-alkynoic acids is time- and concentration-dependent. (b) The 4-carbon acid has a Ki 2 orders of magnitude stronger than the 5-carbon acid, clearly demonstrating the substrate specificity of PDHc. (c) The rate of inactivation of PDH complexes and of resolved E1 by 2-oxo-3-alkynoic acids is enhanced by the addition of ThDP and MgCl2. (d) Pyruvate completely protects E1 and partially protects PDHc from inactivation by 2-oxo-3-butynoic acid. (e) E1 but not E2-E3 is the target of inactivation by 2-oxo-3-butynoic acid. (f) Inactivation of E1 by 2-oxo-3-butynoic acid is accompanied by modification of 1.3 cysteines/E1 monomer. The order of reactivity with the 4-carbon acid was PDHc > POX > PDC. While the order of reactivity with PDHc and POX was 2-oxo-3-butynoic acid > 2-oxo-3-pentynoic acid > 2-oxo-3-heptynoic acid, the order of reactivity was reversed with PDC. |
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Authors:
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A Brown; N Nemeria; J Yi; D Zhang; W B Jordan; R S Machado; J R Guest; F Jordan |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Biochemistry Volume: 36 ISSN: 0006-2960 ISO Abbreviation: Biochemistry Publication Date: 1997 Jul |
Date Detail:
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Created Date: 1997-07-21 Completed Date: 1997-07-21 Revised Date: 2007-11-14 |
Medline Journal Info:
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Nlm Unique ID: 0370623 Medline TA: Biochemistry Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 8071-81 Citation Subset: IM |
Affiliation:
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Department of Chemistry, Rutgers, The State University of New Jersey, Newark, New Jersey 07102, USA. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Bacterial Proteins
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metabolism Enzyme Activation Escherichia coli / enzymology Fatty Acids, Unsaturated / pharmacology* Fungal Proteins / metabolism Kinetics Lactobacillus / enzymology Pyruvate Decarboxylase / metabolism* Pyruvate Dehydrogenase Complex / antagonists & inhibitors*, biosynthesis, metabolism* Pyruvate Oxidase / metabolism* Saccharomyces cerevisiae / enzymology Thiamine Pyrophosphate / metabolism* |
| Grant Support | |
ID/Acronym/Agency:
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GM-50380/GM/NIGMS NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Bacterial Proteins; 0/Fatty Acids, Unsaturated; 0/Fungal Proteins; 0/Pyruvate Dehydrogenase Complex; 154-87-0/Thiamine Pyrophosphate; 56842-75-2/2-keto-3-butynoic acid; EC 1.2.3.3/Pyruvate Oxidase; EC 4.1.1.1/Pyruvate Decarboxylase |
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