Creation of a recombinant peptide substrate for fluorescence resonance energy transfer-based protease assays
| Autor: | Heather L. Lawson, Lin Zhang, Vallathucherry C. Harish, Jason D. Huff, John Owen, Mary Ann Knovich |
|---|---|
| Rok vydání: | 2006 |
| Předmět: |
medicine.medical_treatment
Molecular Sequence Data Biophysics Biochemistry law.invention Substrate Specificity law medicine Fluorescence Resonance Energy Transfer Molecule Amino Acid Sequence Molecular Biology Peptide sequence DNA Primers Protease Base Sequence Chemistry Mutagenesis Substrate (chemistry) Cell Biology Acceptor Recombinant Proteins Förster resonance energy transfer Recombinant DNA Mutagenesis Site-Directed Peptide Hydrolases |
| Zdroj: | Analytical biochemistry. 358(2) |
| ISSN: | 0003-2697 |
| Popis: | based assays. Generation of pure, reliable,high-quality substrate is essential to such assays. SmallFRET substrates can be easily prepared by chemical pro-duction. The preparation of Xuorogenic substrate becomescumbersome when a larger peptide substrate is required orwhen the amino acid residues that Xank the protease cleav-age site are critical to enzyme activity. We sought a novelmethod to generate a large FRET peptide substrate usingfacile DNA recombinant technology.FRET is a phenomenon that describes the alteration inXuorescence emission that occurs with the interaction oftwo molecules in close proximity. Energy transferred froman excited donor molecule (Xuor) to an acceptor molecule(quencher) is dependent on the Forster radius ( |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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