title: Fluorogenic Substrates for In Situ Monitoring of Caspase-3 Activity in Live Cells.
abstract: The in situ detection of caspase-3 activity has applications in the imaging and monitoring of multiple pathologies, notably cancer. A series of cell penetrating FRET-based fluorogenic substrates were designed and synthesised for the detection of caspase-3 in live cells. A variety of modifications of the classical caspase-3 and caspase-7 substrate sequence Asp-Glu-Val-Asp were carried out in order to increase caspase-3 affinity and eliminate caspase-7 cross-reactivity. To allow cellular uptake and good solubility, the substrates were conjugated to a cationic peptoid. The most selective fluorogenic substrate 27, FAM-Ahx-Asp-Leu-Pro-Asp-Lys(MR)-Ahx, conjugated to the cell penetrating peptoid at the C-terminus, was able to detect and quantify caspase-3 activity in apoptotic cells without cross-reactivity by caspase-7.
authors.affiliation: School of Chemistry, EaStCHEM, University of Edinburgh, Joseph Black building, West Mains Road, Edinburgh EH9 3FJ, United Kingdom.
authors.forename: Ana M
authors.initials: AM
authors.lastname: Pérez-López
authors.affiliation: Department of Medicinal and Organic Chemistry, University of Granada, School of Pharmacy, Campus Cartuja s/n - 18071, Granada, Spain.
authors.forename: M Lourdes
authors.initials: ML
authors.lastname: Soria-Gila
authors.affiliation: School of Chemistry, EaStCHEM, University of Edinburgh, Joseph Black building, West Mains Road, Edinburgh EH9 3FJ, United Kingdom.
authors.forename: Emma R
authors.initials: ER
authors.lastname: Marsden
authors.affiliation: School of Chemistry, EaStCHEM, University of Edinburgh, Joseph Black building, West Mains Road, Edinburgh EH9 3FJ, United Kingdom.
authors.forename: Annamaria
authors.initials: A
authors.lastname: Lilienkampf
authors.affiliation: School of Chemistry, EaStCHEM, University of Edinburgh, Joseph Black building, West Mains Road, Edinburgh EH9 3FJ, United Kingdom.
authors.forename: Mark
authors.initials: M
authors.lastname: Bradley
mesh_terms: D053148:Caspase 3 / Q000378:metabolism*; D002460:Cell Line; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D007700:Kinetics; D013379:Substrate Specificity
chemical_list: D005456:Fluorescent Dyes; D053148:Caspase 3