title: A Fluorescent-Labeled Phosphono Bisbenzguanidine As an Activity-Based Probe for Matriptase.
abstract: Activity-based probes are compounds that exclusively form covalent bonds with active enzymes. They can be utilized to profile enzyme activities in vivo, to identify target enzymes and to characterize their function. The design of a new activity-based probe for matriptase, a member of the type II transmembrane serine proteases, is based on linker-connected bis-benzguanidines. An amino acid, introduced as linker, bears the coumarin fluorophore. Moreover, an incorporated phosphonate allows for a covalent interaction with the active-site serine. The resulting irreversible mode of action was demonstrated, leading to enzyme inactivation and, simultaneously, to a fluorescence labeling of matriptase. The ten-step synthetic approach to a coumarin-labeled bis-benzguanidine and its evaluation as activity-based probe for matriptase based on in-gel fluorescence and fluorescence HPLC is reported. HPLC fluorescence detection as a new application for activity-based probes for proteases is demonstrated herein for the first time.
authors.affiliation: Pharmaceutical Institute, Pharmaceutical Chemistry I, University of Bonn, An der Immenburg 4, 53121, Bonn, Germany.
authors.forename: Daniela
authors.initials: D
authors.lastname: Häußler
authors.affiliation: Pharmaceutical Institute, Pharmaceutical Chemistry I, University of Bonn, An der Immenburg 4, 53121, Bonn, Germany.
authors.forename: Anna-Christina
authors.initials: AC
authors.lastname: Schulz-Fincke
authors.affiliation: Pharmaceutical Institute, Pharmaceutical Chemistry I, University of Bonn, An der Immenburg 4, 53121, Bonn, Germany.
authors.forename: Anna-Madeleine
authors.initials: AM
authors.lastname: Beckmann
authors.affiliation: Institute of Pharmaceutical Chemistry, Philipps University of Marburg, Marbacher Weg 6, 35032, Marburg, Germany.
authors.forename: Aline
authors.initials: A
authors.lastname: Keils
authors.affiliation: Pharmaceutical Institute, Pharmaceutical Chemistry I, University of Bonn, An der Immenburg 4, 53121, Bonn, Germany.|Department of Life Science Informatics, B-IT, LIMES Program Unit Chemical Biology and Medicinal Chemistry, University of Bonn, Dahlmannstr. 2, 53113, Bonn, Germany.
authors.forename: Erik
authors.initials: E
authors.lastname: Gilberg
authors.affiliation: Pharmaceutical Institute, Pharmaceutical Chemistry I, University of Bonn, An der Immenburg 4, 53121, Bonn, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Mangold
authors.affiliation: Department of Life Science Informatics, B-IT, LIMES Program Unit Chemical Biology and Medicinal Chemistry, University of Bonn, Dahlmannstr. 2, 53113, Bonn, Germany.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Bajorath
authors.affiliation: Pharmaceutical Institute, Pharmaceutical Chemistry I, University of Bonn, An der Immenburg 4, 53121, Bonn, Germany.
authors.forename: Marit
authors.initials: M
authors.lastname: Stirnberg
authors.affiliation: Institute of Pharmaceutical Chemistry, Philipps University of Marburg, Marbacher Weg 6, 35032, Marburg, Germany.
authors.forename: Torsten
authors.initials: T
authors.lastname: Steinmetzer
authors.affiliation: Pharmaceutical Institute, Pharmaceutical Chemistry I, University of Bonn, An der Immenburg 4, 53121, Bonn, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Gütschow
mesh_terms: D020134:Catalytic Domain; D005456:Fluorescent Dyes / Q000737:chemistry*; D012697:Serine Endopeptidases / Q000737:chemistry* / Q000378:metabolism; D057057:Serine Proteases / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D005456:Fluorescent Dyes; D057057:Serine Proteases; D012697:Serine Endopeptidases; C485595:matriptase
keywords: activity-based probes; fluorescence; matriptase; peptidomimetics; phosphonates