FRET assay for <i>in vitro</i> enzymatic activity of SrtA<sub>ΔN40</sub>.
- Francesca Necchi (342677)
- Vincenzo Nardi-Dei (312280)
- Massimiliano Biagini (247421)
- Michael Assfalg (342678)
- Annalisa Nuccitelli (342679)
- Roberta Cozzi (120732)
- Nathalie Norais (159651)
- John L. Telford (120748)
- C. Daniela Rinaudo (120751)
- Guido Grandi (120744)
- Domenico Maione (120753)
Publication date
February 2013
DOIAbstract
<p>(A) Hydrolytic activity of 3 µM SrtA<sub>ΔN40</sub> with 192 µM fluorogenic peptide Dabcyl-SF<b>LPKTG</b>M-Edans in HEPES buffer in which H<sub>2</sub>O is the only nucleophile. (B) SrtA<sub>ΔN40</sub> (3 µM) catalyzes an <i>in vitro</i> reaction in the presence of 75 mM NH<sub>2</sub>-Gly<sub>3</sub> and 256 µM of fluorogenic peptide Dabcyl-SF<b>LPKTG</b>M-Edans. The reaction rate is equivalent with or without 5 mM calcium ions.</p
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