Thioamides as fluorescence quenching probes: minimalist chromophores to monitor protein dynamics.
J Am Chem Soc
; 132(42): 14718-20, 2010 Oct 27.
Article
em En
| MEDLINE
| ID: mdl-20886849
ABSTRACT
Decreasing the size of spectroscopic probes can afford higher-resolution structural information from fluorescence experiments. Therefore, we have developed p-cyanophenylalanine (Cnf) and backbone thioamides as a fluorophore/quencher pair. Through the examination of a series of thiopeptides, we have determined the working distance for this pair to be 8-30 Å. We have also carried out a proof-of-principle protein-folding experiment in which a Cnf/thioamide-labeled version of villin headpiece HP35 was thermally unfolded while the Cnf/thioamide distance was monitored by fluorescence. For a given protein, thioamide substitutions could be used to track motions with a much greater number of measurements than for current fluorescence probes, providing a dense array of data with which to model conformational changes.
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1
Base de dados:
MEDLINE
Assunto principal:
Tioamidas
/
Proteínas
/
Corantes Fluorescentes
Idioma:
En
Ano de publicação:
2010
Tipo de documento:
Article