Structure, Function, and Mechanism of the Phenylacetate Pathway Hot Dog-fold Thioesterase PaaI
| Autor: | Ryan Kniewel, Lorenzo I. Finci, Christopher D. Lima, Veronica Solorzano, John A. Buglino, Debra Dunaway-Mariano, Jin Wu, Zhihao Zhuang, Feng Song |
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| Rok vydání: | 2006 |
| Předmět: |
Models
Molecular Protein Folding Protein Conformation Stereochemistry Coenzyme A Static Electricity Crystallography X-Ray Biochemistry Substrate Specificity Structure-Activity Relationship chemistry.chemical_compound Protein structure Thioesterase Catalytic Domain Hydrolase Escherichia coli Structure–activity relationship Binding site Protein Structure Quaternary Molecular Biology Phenylacetates chemistry.chemical_classification Binding Sites Escherichia coli Proteins Hydrolysis Temperature DNA Cell Biology Hydrogen-Ion Concentration Protein Structure Tertiary Kinetics Phenylacetate Enzyme Models Chemical Palmitoyl-CoA Hydrolase chemistry Mutation Mutagenesis Site-Directed |
| Zdroj: | Journal of Biological Chemistry. 281:11028-11038 |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.m513896200 |
| Popis: | The structure and biochemical function of the hot dog-fold thioesterase PaaI operative in the aerobic phenylacetate degradation pathway are examined. PaaI showed modest activity with phenylacetyl-coenzyme A, suggestive of a role in coenzyme A release from this pathway intermediate in the event of limiting downstream pathway enzymes. Minimal activity was observed with aliphatic acyl-coenzyme A thioesters, which ruled out PaaI function in the lower phenylacetate pathway. PaaI was most active with ring-hydroxylated phenylacetyl-coenzyme A thioesters. The x-ray crystal structure of the Escherichia coli thioesterase is reported and analyzed to define the structural basis of substrate recognition and catalysis. The contributions of catalytic and substrate binding residues, thus, identified were examined through steady-state kinetic analysis of site-directed mutant proteins. |
| Databáze: | OpenAIRE |
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