Site-specific interaction of thrombin and inhibitors observed by fluorescence correlation spectroscopy.
AUTOR(ES)
Klingler, J
RESUMO
We report on the application of fluorescence correlation spectroscopy (FCS) to observe the interaction between thrombin and thrombin inhibitors. Two site-specific fluorescent labels were used to distinguish between inhibitors directed to the active site, the exosite, or both binding sites of thrombin. For several well-known inhibitors of thrombin, the binding sites observed by FCS correspond to previous studies. The interaction of the recently discovered thrombin inhibitor ornithodorin from the tick Ornithodorus moubata with thrombin was investigated. It was found that this inhibitor, like hirudin and rhodniin, binds to both the active site and exosite of thrombin simultaneously. This study shows the feasibility of FCS as a sensitive and selective method for observing protein-ligand interactions. As an additional technique, simultaneous labeling with both fluorescent labels was successfully demonstrated.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=1181121Documentos Relacionados
- Site-specific interaction of DNA gyrase with DNA.
- Site-specific labeling of the ribosome for single-molecule spectroscopy
- DNA binding and oligomerization of NtrC studied by fluorescence anisotropy and fluorescence correlation spectroscopy.
- Ultrasensitive hybridization analysis using fluorescence correlation spectroscopy.
- Isolation and analysis of inhibitors of transposon Tn3 site-specific recombination.