Development of the first internally-quenched fluorescent substrates of human cathepsin C

The application in the enzyme detection in biological samples

Monika Łęgowska, Yveline Hamon, Anna Wojtysiak, Renata Grzywa, Marcin Sieńczyk, Timo Burster, Brice Korkmaz, Adam Lesner

Research output: Contribution to journalArticle

2 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)91-102
Number of pages12
JournalArchives of Biochemistry and Biophysics
Volume612
DOIs
Publication statusPublished - Dec 15 2016
Externally publishedYes

Fingerprint

Cathepsin C
Substrates
Enzymes
Exopeptidases
Fluorescence Resonance Energy Transfer
Fluids
T-cells
Bronchoalveolar Lavage Fluid
Serine Proteases
Cytotoxic T-Lymphocytes
Human Activities
Mast Cells
Cysteine
Catalytic Domain
Neutrophils
Chemical activation
Urine
Peptides

Keywords

  • Catalytic Domain
  • Cathepsin C
  • Cathepsin L
  • Fluorescence Resonance Energy Transfer
  • Humans
  • Hydrogen-Ion Concentration
  • Inflammation
  • Kinetics
  • Mast Cells
  • Microscopy, Fluorescence
  • Molecular Conformation
  • Molecular Docking Simulation
  • Neutrophils
  • Peptides
  • Protein Binding
  • Recombinant Proteins
  • Substrate Specificity
  • T-Lymphocytes, Cytotoxic
  • Journal Article

Cite this

Development of the first internally-quenched fluorescent substrates of human cathepsin C : The application in the enzyme detection in biological samples. / Łęgowska, Monika; Hamon, Yveline; Wojtysiak, Anna; Grzywa, Renata; Sieńczyk, Marcin; Burster, Timo; Korkmaz, Brice; Lesner, Adam.

In: Archives of Biochemistry and Biophysics, Vol. 612, 15.12.2016, p. 91-102.

Research output: Contribution to journalArticle

Łęgowska, Monika ; Hamon, Yveline ; Wojtysiak, Anna ; Grzywa, Renata ; Sieńczyk, Marcin ; Burster, Timo ; Korkmaz, Brice ; Lesner, Adam. / Development of the first internally-quenched fluorescent substrates of human cathepsin C : The application in the enzyme detection in biological samples. In: Archives of Biochemistry and Biophysics. 2016 ; Vol. 612. pp. 91-102.
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AU - Grzywa, Renata

AU - Sieńczyk, Marcin

AU - Burster, Timo

AU - Korkmaz, Brice

AU - Lesner, Adam

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