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

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.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-27AB-D Version Permalink: http://hdl.handle.net/11858/00-001M-0000-002E-75F9-5
Genre: Journal Article
Alternative Title : Intern. J. Environ. Anal. Chem.

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IntJEnvironmentalAnalytChem_85_2005_741.pdf (Any fulltext), 438KB
 
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 Creators:
Marmé, Nicole, Author
Weston, Kenneth D., Author
Staudt, Thorsten, Author
Spatz, Joachim P.1, 2, Author              
Knemeyer, Jens-Peter, Author
Affiliations:
1Cellular Biophysics, Max Planck Institute for Medical Research, Max Planck Society, ou_2364731              
2Biophysical Chemistry, Institute of Physical Chemistry, University of Heidelberg, 69120 Heidelberg, Germany, ou_persistent22              

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Free keywords: Exopeptidase-assay; Self-quenching substrates; Single-molecule imaging
 Abstract: 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.

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Language(s): eng - English
 Dates: 2004-10-042005-03-112007-01-252005
 Publication Status: Published in print
 Pages: 11
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: eDoc: 240004
DOI: 10.1080/03067310500147720
 Degree: -

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Title: International Journal of Environmental Analytical Chemistry
Source Genre: Journal
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Publ. Info: New York : Gordon and Breach.
Pages: - Volume / Issue: 85 (9-11) Sequence Number: - Start / End Page: 741 - 751 Identifier: ISSN: 0306-7319
CoNE: https://pure.mpg.de/cone/journals/resource/954925515508