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Development of Chemical Tools to Monitor Human Kallikrein 13 (KLK13) Activity.

Natalia GrubaEwa BieleckaMagdalena WysockaAnna WojtysiakMagdalena Brzezińska-BodalKamila SychowskaMagdalena KalińskaMałgorzata MagochAleksandra PęcakKatherine FalkowskiMagdalena WiśniewskaLaura SąsiadekKarolina PłazaEileen KrollAnastasija PejkovskaMaren RehdersKlaudia BrixGrzegorz DubinTomasz KantykaJan PotempaAdam Lesner
Published in: International journal of molecular sciences (2019)
Kallikrein 13 (KLK13) was first identified as an enzyme that is downregulated in a subset of breast tumors. This serine protease has since been implicated in a number of pathological processes including ovarian, lung and gastric cancers. Here we report the design, synthesis and deconvolution of libraries of internally quenched fluorogenic peptide substrates to determine the specificity of substrate binding subsites of KLK13 in prime and non-prime regions (according to the Schechter and Berger convention). The substrate with the consensus sequential motive ABZ-Val-Arg-Phe-Arg-ANB-NH₂ demonstrated selectivity towards KLK13 and was successfully converted into an activity-based probe by the incorporation of a chloromethylketone warhead and biotin bait. The compounds described may serve as suitable tools to detect KLK13 activity in diverse biological samples, as exemplified by overexpression experiments and targeted labeling of KLK13 in cell lysates and saliva. In addition, we describe the development of selective activity-based probes targeting KLK13, to our knowledge the first tool to analyze the presence of the active enzyme in biological samples.
Keyphrases
  • endothelial cells
  • healthcare
  • cancer therapy
  • cell proliferation
  • small molecule
  • stem cells
  • cell therapy
  • single cell
  • bone marrow
  • transcription factor
  • fluorescence imaging
  • binding protein