New fluorescent and photoactivable analogs acting on nucleotide binding enzymes
✍ Scribed by Robert S. Sarfati; Abdelkader Namane; Hélène Munier; Octavian Bârzu
- Book ID
- 104225233
- Publisher
- Elsevier Science
- Year
- 1991
- Tongue
- French
- Weight
- 254 KB
- Volume
- 32
- Category
- Article
- ISSN
- 0040-4039
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✦ Synopsis
We describe a seven-step synthesis of 8-azido-3'O-anthraniloyk2ilADP and 2'dATP from 8-azido-2 'deoxyadenosine. These compounds can be used asji'ttorescent and photoactivable probesfor the nucleotide-binding site of kinases or cyclases.
Nucleotide analogs with functional reactive groups have been increasingly used to characterize substrate and/or effector sites of ATP-dependent enzymes -1 3. The most widely employed compounds are the azidoderivatives of ATP or ADP. Under weak ultraviolet light, the azido group changes rapidly to the corresponding nitrene which can then react with the side chain of several amino acid residues4-9.
Recently, we have shown that the fluorescent 3'-0-anthraniloyl derivative of 2'dATP (Ant-dATP) is a strong competitive inhibitor (Ki 10 PM) of two calmodulin (CaM)-activated bacterial adenylate cyclaseslO.
Binding of this analog to Bordetella pertussis or Bacillus anthracis enzyme is specific and is accompanied by a several-fold increase in its quantum yield. As 8-azido-ATP showed no or little decrease in its affinity for the active site of adenylate cyclase11112, we predicted that 8-azido-3'-0-anthraniloyl-2'dATP might act as a useful fluorescent and photoactivable probe for the nucleotide-binding site of bacterial adenylate cyclase(s) with approximately ten times higher affinity than that of the natural substrate.