title: Design of a highly selective quenched activity-based probe and its application in dual color imaging studies of cathepsin S activity localization.
abstract: The cysteine cathepsins are a group of 11 proteases whose function was originally believed to be the degradation of endocytosed material with a high degree of redundancy. However, it has become clear that these enzymes are also important regulators of both health and disease. Thus, selective tools that can discriminate between members of this highly related class of enzymes will be critical to further delineate the unique biological functions of individual cathepsins. Here we present the design and synthesis of a near-infrared quenched activity-based probe (qABP) that selectively targets cathepsin S which is highly expressed in immune cells. Importantly, this high degree of selectivity is retained both in vitro and in vivo. In combination with a new green-fluorescent pan-reactive cysteine cathepsin qABP we performed dual color labeling studies in bone marrow-derived immune cells and identified vesicles containing exclusively cathepsin S activity. This observation demonstrates the value of our complementary cathepsin probes and provides evidence for the existence of specific localization of cathepsin S activity in dendritic cells.
authors.forename: Kristina
authors.initials: K
authors.lastname: Oresic Bender
authors.forename: Leslie
authors.initials: L
authors.lastname: Ofori
authors.forename: Wouter A
authors.initials: WA
authors.lastname: van der Linden
authors.forename: Elliot D
authors.initials: ED
authors.lastname: Mock
authors.affiliation: ∥Department of Chemistry, University of California-Berkeley, Berkeley, California 94720, United States.
authors.forename: Gopal K
authors.initials: GK
authors.lastname: Datta
authors.affiliation: ∥Department of Chemistry, University of California-Berkeley, Berkeley, California 94720, United States.
authors.forename: Somenath
authors.initials: S
authors.lastname: Chowdhury
authors.forename: Hao
authors.initials: H
authors.lastname: Li
authors.forename: Ehud
authors.initials: E
authors.lastname: Segal
authors.forename: Mateo
authors.initials: M
authors.lastname: Sanchez Lopez
authors.affiliation: ∥Department of Chemistry, University of California-Berkeley, Berkeley, California 94720, United States.
authors.forename: Jonathan A
authors.initials: JA
authors.lastname: Ellman
authors.affiliation: ⊥Department of Tumor Immunology, Radboud University Medical Center, Radboud Institute for Molecular Life Sciences, 6500 HB Nijmegen, The Netherlands.
authors.forename: Carl G
authors.initials: CG
authors.lastname: Figdor
authors.forename: Matthew
authors.initials: M
authors.lastname: Bogyo
authors.affiliation: ⊥Department of Tumor Immunology, Radboud University Medical Center, Radboud Institute for Molecular Life Sciences, 6500 HB Nijmegen, The Netherlands.
authors.forename: Martijn
authors.initials: M
authors.lastname: Verdoes
mesh_terms: D000818:Animals; D002403:Cathepsins / Q000737:chemistry* / Q000378:metabolism*; D003116:Color; D003713:Dendritic Cells / Q000201:enzymology; D015195:Drug Design; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D007259:Infrared Rays; D008325:Mammary Neoplasms, Experimental / Q000201:enzymology; D051379:Mice; D061848:Optical Imaging / Q000379:methods*; D000067996:RAW 264.7 Cells; D013379:Substrate Specificity
chemical_list: D005456:Fluorescent Dyes; D002403:Cathepsins; C028217:cathepsin S