Synthesis and evaluation of a selective fluorogenic Pup derived assay reagent for Dop, a potential drug target in Mycobacterium tuberculosis

Chembiochem. 2012 Sep 24;13(14):2056-60. doi: 10.1002/cbic.201200460. Epub 2012 Aug 24.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amidohydrolases / antagonists & inhibitors
  • Amidohydrolases / metabolism*
  • Amino Acid Sequence
  • Bacterial Proteins / antagonists & inhibitors
  • Bacterial Proteins / metabolism*
  • Biocatalysis
  • Enzyme Inhibitors / chemical synthesis
  • Enzyme Inhibitors / chemistry
  • Fluorescent Dyes / chemistry*
  • High-Throughput Screening Assays
  • Hydrolysis
  • Kinetics
  • Molecular Sequence Data
  • Mycobacterium tuberculosis / drug effects*
  • Peptides / chemical synthesis
  • Peptides / chemistry
  • Peptides / metabolism
  • Substrate Specificity
  • Ubiquitins / antagonists & inhibitors
  • Ubiquitins / metabolism*
  • Virulence Factors / antagonists & inhibitors
  • Virulence Factors / metabolism*

Substances

  • Bacterial Proteins
  • Enzyme Inhibitors
  • Fluorescent Dyes
  • Peptides
  • Pup protein, Mycobacterium tuberculosis
  • Ubiquitins
  • Virulence Factors
  • Amidohydrolases
  • Dop protein, Mycobacterium tuberculosis