Structure of a ricin mutant showing rescue of activity by a noncatalytic residue
| Autor: | Jon D. Robertus, Youngsoo Kim, Art Frankel, Debra Mlsna, Arthur F. Monzingo, Michael P. Ready |
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| Rok vydání: | 1992 |
| Předmět: |
Stereochemistry
Mutant Glutamic Acid Ricin Biochemistry Ribosome Catalysis Structure-Activity Relationship chemistry.chemical_compound Glutamates Mutant protein Ribose Escherichia coli Animals Amino Acid Sequence Carboxylate Enzyme kinetics Binding Sites biology Active site Kinetics chemistry Mutation biology.protein Artemia |
| Zdroj: | Biochemistry. 31:3294-3296 |
| ISSN: | 1520-4995 0006-2960 |
| Popis: | Ricin A chain is an N-glycosidase which removes a single adenine base from a conservative loop of 28S rRNA, thereby inactivating eukaryotic ribosomes. The mechanism of action has been proposed to include transition-state stabilization of an oxycarbonium ion on the substrate ribose by interaction with Glu 177. Conversion of Glu 177 to Gln reduces activity nearly 200-fold [Ready, M. P., Kim, Y., & Robertus, J. D. (1991) Proteins: Struct., Funct., Genet. 10, 270-278] while conversion to Ala (E177A) reduces activity only 20-fold [Schlossman, D., Withers, D., Welsh, P., Alexander, A., Robertus, J., & Frankel, A. (1989) Mol. Cell. Biol. 9, 5012-5021]. X-ray analysis of the latter mutant protein shows that a residue at the edge of the active site, Glu 208, rotates into the space left vacant by the mutation. Its rearranged carboxylate partially substitutes for that of Glu 177. This is equivalent to the rescue of enzyme activity by a second-site reversion. Kinetic analysis shows the E177A mutation affects kcat and not Km, consistent with the notion that the carboxylate serves in transition-state stabilization. |
| Databáze: | OpenAIRE |
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