Nucleoside binding site of herpes simplex type 1 thymidine kinase analyzed by X-ray crystallography
| Autor: | J, Vogt, R, Perozzo, A, Pautsch, A, Prota, P, Schelling, B, Pilger, G, Folkers, L, Scapozza, G E, Schulz |
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| Rok vydání: | 2000 |
| Předmět: |
Adenosine
Binding Sites Adenine Organophosphonates Water Nucleosides Stereoisomerism Herpesvirus 1 Human Crystallography X-Ray Antiviral Agents Thymidine Kinase Adenosine Monophosphate Protein Structure Tertiary Substrate Specificity Viral Proteins Amino Acid Substitution Catalytic Domain Mutation Prodrugs Enzyme Inhibitors Thymidine |
| Zdroj: | Proteins. 41(4) |
| ISSN: | 0887-3585 |
| Popis: | The crystal structures of the full-length Herpes simplex virus type 1 thymidine kinase in its unligated form and in a complex with an adenine analogue have been determined at 1.9 A resolution. The unligated enzyme contains four water molecules in the thymidine pocket and reveals a small induced fit on substrate binding. The structure of the ligated enzyme shows for the first time a bound adenine analogue after numerous complexes with thymine and guanine analogues have been reported. The adenine analogue constitutes a new lead compound for enzyme-prodrug gene therapy. In addition, the structure of mutant Q125N modifying the binding site of the natural substrate thymidine in complex with this substrate has been established at 2.5 A resolution. It reveals that neither the binding mode of thymidine nor the polypeptide backbone conformation is altered, except that the two major hydrogen bonds to thymidine are replaced by a single water-mediated hydrogen bond, which improves the relative acceptance of the prodrugs aciclovir and ganciclovir compared with the natural substrate. Accordingly, the mutant structure represents a first step toward improving the virus-directed enzyme-prodrug gene therapy by enzyme engineering. |
| Databáze: | OpenAIRE |
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