title: Design of Selective Substrates and Activity-Based Probes for Hydrolase Important for Pathogenesis 1 (HIP1) from Mycobacterium tuberculosis.
abstract: Although serine proteases are important mediators of Mycobacterium tuberculosis (Mtb) virulence, there are currently no tools to selectively block or visualize members of this family of enzymes. Selective reporter substrates or activity-based probes (ABPs) could provide a means to monitor infection and response to therapy using imaging methods. Here, we use a combination of substrate selectivity profiling and focused screening to identify optimized reporter substrates and ABPs for the Mtb "Hydrolase important for pathogenesis 1" (Hip1) serine protease. Hip1 is a cell-envelope-associated enzyme with minimal homology to host proteases, making it an ideal target for probe development. We identified substituted 7-amino-4-chloro-3-(2-bromoethoxy)isocoumarins as irreversible inhibitor scaffolds. Furthermore, we used specificity data to generate selective reporter substrates and to further optimize a selective chloroisocoumarin inhibitor. These new reagents are potentially useful in delineating the roles of Hip1 during pathogenesis or as diagnostic imaging tools for specifically monitoring Mtb infections.
authors.forename: Christian S
authors.initials: CS
authors.lastname: Lentz
authors.forename: Alvaro A
authors.initials: AA
authors.lastname: Ordonez
authors.affiliation: Division of Bioorganic Chemistry, Faculty of Chemistry, Wroclaw University of Technology , Wybrzeze Wyspianskiego 27, 50-370 Wroclaw, Poland.
authors.forename: Paulina
authors.initials: P
authors.lastname: Kasperkiewicz
authors.affiliation: Department of Pharmaceutical Chemistry, University of California-San Francisco , San Francisco, United States.
authors.forename: Florencia
authors.initials: F
authors.lastname: La Greca
authors.affiliation: Department of Pharmaceutical Chemistry, University of California-San Francisco , San Francisco, United States.|Skaggs School of Pharmacy and Pharmaceutical Sciences, University of California-San Diego , La Jolla, California 92093 United States.
authors.forename: Anthony J
authors.initials: AJ
authors.lastname: O'Donoghue
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Schulze
authors.affiliation: Department of Chemistry, Georgia Institute of Technology , Atlanta, Georgia 30332, United States.
authors.forename: James C
authors.initials: JC
authors.lastname: Powers
authors.affiliation: Department of Pharmaceutical Chemistry, University of California-San Francisco , San Francisco, United States.
authors.forename: Charles S
authors.initials: CS
authors.lastname: Craik
authors.affiliation: Division of Bioorganic Chemistry, Faculty of Chemistry, Wroclaw University of Technology , Wybrzeze Wyspianskiego 27, 50-370 Wroclaw, Poland.
authors.forename: Marcin
authors.initials: M
authors.lastname: Drag
authors.forename: Sanjay K
authors.initials: SK
authors.lastname: Jain
authors.forename: Matthew
authors.initials: M
authors.lastname: Bogyo
mesh_terms: D000818:Animals; D000995:Antitubercular Agents / Q000737:chemistry* / Q000494:pharmacology; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015195:Drug Design; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D015394:Molecular Structure; D009169:Mycobacterium tuberculosis / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D057057:Serine Proteases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity; D014376:Tuberculosis / Q000382:microbiology*
chemical_list: D000995:Antitubercular Agents; D001426:Bacterial Proteins; D004791:Enzyme Inhibitors; C000598517:Hip1 protein, Mycobacterium tuberculosis; D057057:Serine Proteases
keywords: Mycobacterium tuberculosis; activity-based probe; fluorogenic substrate; hydrolase important for pathogenesis 1 (Hip1); protease; substrate profiling