title: Development of the first internally-quenched fluorescent substrates of human cathepsin C: The application in the enzyme detection in biological samples.
abstract: Cathepsin C is a widely expressed cysteine exopeptidase that is mostly recognized for the activation of the granule-associated proinflammatory serine proteases in neutrophils, cytotoxic T lymphocytes and mast cells. It has been shown that the enzyme can be secreted extracellularly; however, its occurrence in human bodily fluids/physiological samples has not been thoroughly studied. In the course of this study, the first fluorescence resonance energy transfer peptides for the measurement of the activity of human cathepsin C were designed and synthesized. Two series of tetra- and pentapeptide substrates enabled the detailed S' specificity study of cathepsin C, which has been examined for the first time. The extensive enzymatic studies of the obtained compounds resulted in the selection of the highly specific and selective substrate Thi-Ala(Mca)-Ser-Gly-Tyr(3-NO2)-NH2, which was successfully employed for the detection of cathepsin C activity in complex biological samples such as cell lysates, urine and bronchoalveolar lavage fluids. Molecular docking of the selected substrate was performed in order to better understand the binding mode of the substrates in the active site of cathepsin C.
authors.affiliation: Faculty of Chemistry, University of Gdansk, 80-308, Gdansk, Poland.
authors.forename: Monika
authors.initials: M
authors.lastname: Łęgowska
authors.affiliation: INSERM U-1100, Centre d'Etude des Pathologies Respiratoires, Université François Rabelais, 37032, Tours, France.
authors.forename: Yveline
authors.initials: Y
authors.lastname: Hamon
authors.affiliation: Faculty of Chemistry, University of Gdansk, 80-308, Gdansk, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Wojtysiak
authors.affiliation: Faculty of Chemistry, Wroclaw University of Science and Technology, 50-370, Wroclaw, Poland.
authors.forename: Renata
authors.initials: R
authors.lastname: Grzywa
authors.affiliation: Faculty of Chemistry, Wroclaw University of Science and Technology, 50-370, Wroclaw, Poland.
authors.forename: Marcin
authors.initials: M
authors.lastname: Sieńczyk
authors.affiliation: Department of Neurosurgery, Ulm University Medical Centre, 89081, Ulm, Germany.
authors.forename: Timo
authors.initials: T
authors.lastname: Burster
authors.affiliation: INSERM U-1100, Centre d'Etude des Pathologies Respiratoires, Université François Rabelais, 37032, Tours, France.
authors.forename: Brice
authors.initials: B
authors.lastname: Korkmaz
authors.affiliation: Faculty of Chemistry, University of Gdansk, 80-308, Gdansk, Poland. Electronic address: adam.lesner@ug.edu.pl.
authors.forename: Adam
authors.initials: A
authors.lastname: Lesner
mesh_terms: D020134:Catalytic Domain; D020789:Cathepsin C / Q000737:chemistry*; D056668:Cathepsin L / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007249:Inflammation; D007700:Kinetics; D008407:Mast Cells / Q000166:cytology; D008856:Microscopy, Fluorescence / Q000379:methods*; D008968:Molecular Conformation; D062105:Molecular Docking Simulation; D009504:Neutrophils / Q000378:metabolism; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry; D013379:Substrate Specificity; D013602:T-Lymphocytes, Cytotoxic / Q000166:cytology
chemical_list: D010455:Peptides; D011994:Recombinant Proteins; D020789:Cathepsin C; D056668:Cathepsin L
keywords: Cathepsin C; FRET substrates; Peptidomimetics; cysteine proteases