Eliminating caspase-7 and cathepsin B cross-reactivity on fluorogenic caspase-3 substrates

Mol Biosyst. 2016 Mar;12(3):693-6. doi: 10.1039/c5mb00730e.

Abstract

11 FRET-based fluorogenic substrates were constructed using the pentapeptide template Asp-Glu-X2-Asp-X1', and evaluated with caspase-3, caspase-7 and cathepsin B. The sequence Asp-Glu-Pro-Asp-Ser was able to selectively quantify caspase-3 activity in vitro without notable caspase-7 and cathepsin B cross-reactivity, while exhibiting low μM KM values and good catalytic efficiencies (7.0-16.9 μM(-1) min(-1)).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Caspase 3 / metabolism*
  • Caspase 7 / metabolism*
  • Cathepsin B / metabolism*
  • Fluorescence Resonance Energy Transfer
  • Fluorescent Dyes / chemistry
  • Fluorescent Dyes / metabolism*
  • Humans
  • Kinetics
  • Peptides / chemistry
  • Peptides / metabolism
  • Substrate Specificity

Substances

  • Fluorescent Dyes
  • Peptides
  • Caspase 3
  • Caspase 7
  • Cathepsin B