title: Eliminating caspase-7 and cathepsin B cross-reactivity on fluorogenic caspase-3 substrates.
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)).
authors.affiliation: EaStCHEM, School of Chemistry, University of Edinburgh, West Mains Road, EH9 3FJ Edinburgh, UK. annamaria.lilienkampf@ed.ac.uk mark.bradley@ed.ac.uk.
authors.forename: Martha
authors.initials: M
authors.lastname: Mackay
authors.affiliation: EaStCHEM, School of Chemistry, University of Edinburgh, West Mains Road, EH9 3FJ Edinburgh, UK. annamaria.lilienkampf@ed.ac.uk mark.bradley@ed.ac.uk.
authors.forename: Ana M
authors.initials: AM
authors.lastname: Pérez-López
authors.affiliation: EaStCHEM, School of Chemistry, University of Edinburgh, West Mains Road, EH9 3FJ Edinburgh, UK. annamaria.lilienkampf@ed.ac.uk mark.bradley@ed.ac.uk.
authors.forename: Mark
authors.initials: M
authors.lastname: Bradley
authors.affiliation: EaStCHEM, School of Chemistry, University of Edinburgh, West Mains Road, EH9 3FJ Edinburgh, UK. annamaria.lilienkampf@ed.ac.uk mark.bradley@ed.ac.uk.
authors.forename: Annamaria
authors.initials: A
authors.lastname: Lilienkampf
mesh_terms: D053148:Caspase 3 / Q000378:metabolism*; D053179:Caspase 7 / Q000378:metabolism*; D002401:Cathepsin B / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D005456:Fluorescent Dyes; D010455:Peptides; D053148:Caspase 3; D053179:Caspase 7; D002401:Cathepsin B