Overexpression, purification and crystallographic analysis of a unique adenosine kinase fromMycobacterium tuberculosis
| Autor: | Mary C. Long, Rongbao Li, Vincent Escuyer, Yimin Wang, William B. Parker, Senthil Ranganathan |
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| Rok vydání: | 2005 |
| Předmět: |
Adenosine monophosphate
Biophysics Adenosine kinase Biochemistry Mycobacterium tuberculosis chemistry.chemical_compound Adenosine deaminase Bacterial Proteins X-Ray Diffraction Structural Biology Escherichia coli Genetics medicine Cloning Molecular Adenosine Kinase chemistry.chemical_classification biology Condensed Matter Physics biology.organism_classification Adenosine ADK Crystallography Enzyme Mechanism of action chemistry Crystallization Communications biology.protein Volatilization medicine.symptom Crystallization Dimerization medicine.drug |
| Zdroj: | Acta Crystallographica Section F Structural Biology and Crystallization Communications. 61:553-557 |
| ISSN: | 1744-3091 |
| DOI: | 10.1107/s1744309105013473 |
| Popis: | Adenosine kinase from Mycobacterium tuberculosis is the only prokaryotic adenosine kinase that has been isolated and characterized. The enzyme catalyzes the phosphorylation of adenosine to adenosine monophosphate and is involved in the activation of 2-methyladenosine, a compound that has demonstrated selective activity against M. tuberculosis. The mechanism of action of 2-methyladenosine is likely to be different from those of current tuberculosis treatments and this compound (or other adenosine analogs) may prove to be a novel therapeutic intervention for this disease. The M. tuberculosis adenosine kinase was overexpressed in Escherichia coli and the enzyme was purified with activity comparable to that reported previously. The protein was crystallized in the presence of adenosine using the vapour-diffusion method. The crystals diffracted X-rays to high resolution and a complete data set was collected to 2.2 A using synchrotron radiation. The crystal belonged to space group P3(1)21, with unit-cell parameters a = 70.2, c = 111.6 A, and contained a single protein molecule in the asymmetric unit. An initial structural model of the protein was obtained by the molecular-replacement method, which revealed a dimeric structure. The monomers of the dimer were related by twofold crystallographic symmetry. An understanding of how the M. tuberculosis adenosine kinase differs from the human homolog should aid in the design of more potent and selective antimycobacterial agents that are selectively activated by this enzyme. |
| Databáze: | OpenAIRE |
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