title: Synthesis and evaluation of a selective fluorogenic Pup derived assay reagent for Dop, a potential drug target in Mycobacterium tuberculosis.
abstract: A litter of pups: The synthesis and in vitro evaluation of new Pup-based fluorogenic substrates for Dop, the mycobacterial depupylase, are described. A full-length Pup-amidomethylcoumarin conjugate as well as an amino-terminus-truncated analogue exhibited high sensitivity and specificity towards hydrolysis by Dop. The substrates developed here might find application as high-throughput screening assay reagents for the identification of Dop inhibitors.
authors.affiliation: Division of Cell Biology, Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands.
authors.forename: Remco
authors.initials: R
authors.lastname: Merkx
authors.forename: Kristin E
authors.initials: KE
authors.lastname: Burns
authors.forename: Paul
authors.initials: P
authors.lastname: Slobbe
authors.forename: Farid
authors.initials: F
authors.lastname: El Oualid
authors.forename: Dris
authors.initials: D
authors.lastname: El Atmioui
authors.forename: K Heran
authors.initials: KH
authors.lastname: Darwin
authors.forename: Huib
authors.initials: H
authors.lastname: Ovaa
mesh_terms: D000581:Amidohydrolases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D055162:Biocatalysis; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry*; D057166:High-Throughput Screening Assays; D006868:Hydrolysis; D007700:Kinetics; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000187:drug effects*; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism; D013379:Substrate Specificity; D014452:Ubiquitins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D037521:Virulence Factors / Q000037:antagonists & inhibitors / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D010455:Peptides; C533288:Pup protein, Mycobacterium tuberculosis; D014452:Ubiquitins; D037521:Virulence Factors; D000581:Amidohydrolases; C556013:Dop protein, Mycobacterium tuberculosis