| DOI | Resolve DOI: https://doi.org/10.1074/jbc.M412028200 |
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| Author | Search for: Lindner, Holger A.1; Search for: Nadeau, Guy1; Search for: Matte, Allan1; Search for: Michel, Gurvan; Search for: Ménard, Robert1; Search for: Cygler, Miroslaw1 |
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| Name affiliation | - National Research Council of Canada. NRC Biotechnology Research Institute
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| Format | Text, Article |
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| Journal title | The Journal of Biological Chemistry |
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| ISSN | 0021-9258
1083-351X |
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| Volume | 280 |
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| Issue | 8 |
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| Pages | 7162–7169 |
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| Subject | 5'-Guanylic Acid; Escherichia coli; mutagenesis; pharmaceutical |
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| Abstract | YdiB and its paralog AroE are members of the quinate/shikimate 5-dehdrogenase family. Enzymes from this family function in the shikimate pathway that is essential for survival of microorganisms and plants and represent potential drug targets. Recent YdiB and AroE crystal structures revealed the presence of a NAD(P)-binding and a catalytic domain. We carried out site-directed mutagenesis of 8 putative active site residues in YdiB from Escherichia coli and analyzed structural and kinetic properties of the mutant enzymes. Our data indicate critical roles for an invariant lysine and aspartate residue in substrate binding and allowed us to differentiate between two previously proposed models for the binding of the substrate in the active site. Comparison of several YdiB and AroE structures led us to conclude that, upon cofactor binding and domain closure, the 2 identified binding residues are repositioned to bind to the substrate. Although the lysine residue contributes to some extent to the stabilization of the transition state, we did not identify any residue as catalytically essential. This indicates that catalysis does not operate through a general acid-base mechanism, as thought originally. Our improved understanding of the medically and agriculturally important quinate/shikimate 5-dehydrogenase family at the molecular level may prove useful in the development of novel herbicides and antimicrobial agents. |
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| Publication date | 2004-12-13 |
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| Publisher | American Society for Biochemistry and Molecular Biology |
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| Language | English |
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| Peer reviewed | Yes |
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| NRC number | NRC-BRI-46231 |
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| NPARC number | 3539417 |
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| Export citation | Export as RIS |
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| Report a correction | Report a correction |
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| Record identifier | e81eeca8-292d-4c13-a1e6-ecad18feb080 |
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| Record created | 2009-03-01 |
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| Record modified | 2020-04-17 |
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