Deutsch
 
Benutzerhandbuch Datenschutzhinweis Impressum Kontakt
  DetailsucheBrowse

Datensatz

DATENSATZ AKTIONENEXPORT

Freigegeben

Zeitschriftenartikel

A fluorescence-based assay for exopeptidases using self-quenching peptide probes with single-molecule sensitivity

MPG-Autoren
/persons/resource/persons76135

Spatz,  Joachim P.
Cellular Biophysics, Max Planck Institute for Medical Research, Max Planck Society;
Biophysical Chemistry, Institute of Physical Chemistry, University of Heidelberg, 69120 Heidelberg, Germany;

Volltexte (frei zugänglich)
Es sind keine frei zugänglichen Volltexte verfügbar
Ergänzendes Material (frei zugänglich)
Es sind keine frei zugänglichen Ergänzenden Materialien verfügbar
Zitation

Marmé, N., Weston, K. D., Staudt, T., Spatz, J. P., & Knemeyer, J.-P. (2005). A fluorescence-based assay for exopeptidases using self-quenching peptide probes with single-molecule sensitivity. International Journal of Environmental Analytical Chemistry, 85(9-11), 741-751. doi:10.1080/03067310500147720.


Zitierlink: http://hdl.handle.net/11858/00-001M-0000-0010-27AB-D
Zusammenfassung
In recent years, the interest in assaying exopeptidases has become increasingly important because they are significantly involved in many diseases like cancer. To date, no generally applicable fluorescence-based detection method has been developed because commercially available doubly-labeled substrates are not always digested by exopeptidases. In this article we present a new method for the sensitive detection of exopeptidases based on fluorescently-labeled substrates containing only one fluorophore that is efficiently quenched by an adjacent tryptophan residue via photoinduced electron transfer. Because of their well-known properties we chose carboxypeptidase A (CPA) as a model system. The self-quenching probes were used in homogeneous solution as well as on cross-linked PEG-coated surfaces in combination with single-molecule imaging techniques. However, even with standard fluorescence spectrometers we achieved sensitivity below the picomolar range.