Spliced analogues of trypsin inhibitor SFTI-1 and their application for tracing proteolysis and delivery of cargos inside the cells

Magdalena Filipowicz, Natalia Ptaszyńska, Katarzyna Olkiewicz, Dawid Dębowski, Kamila Ćwikłowska, Timo Burster, Michał Pikuła, Adam Krzystyniak, Anna Łęgowska, Krzysztof Rolka

Research output: Contribution to journalArticlepeer-review

4 Citations (Scopus)

Abstract

A series of analogues of trypsin inhibitor SFTI-1 were designed and synthesized to monitor peptide splicing. In the middle part of the SFTI-1 analogues, which is released upon incubation with proteinase, the RGD sequence or an acceptor of fluorescence for FRET was introduced. The results of studies with trypsin confirmed that the designed analogues underwent peptide splicing. Furthermore, we showed that a FRET displaying SFTI-1 analogue was internalized into the HaCaT keratinocytes, where it was degraded. Therefore, both proteolysis and the reduction of the disulfide bridge of the peptide took place. As a result, such analogues are a convenient tool to trace the proteolytic activity inside the cell. However, the cytotoxicity of SFTI-1 analogues grafted with the RGD sequence did not correlate with their susceptibility to peptide splicing. Nevertheless, these peptides were slightly more active than the reference peptide (GRGDNP). Interestingly, one of the analogues assigned as [desSer6 ]VI, under experimental conditions, appeared significantly more cytotoxic towards cancer cells U87-MG in contrast to the reference peptide.

Original languageEnglish
JournalBiopolymers
Volume108
Issue number2
DOIs
Publication statusPublished - Mar 2017
Externally publishedYes

Keywords

  • Amino Acid Sequence
  • Cell Line
  • Cell Line, Tumor
  • Cell Survival
  • Fluorescence Resonance Energy Transfer
  • Humans
  • Keratinocytes
  • Mass Spectrometry
  • Microscopy, Fluorescence
  • Oligopeptides
  • Peptides
  • Proteolysis
  • Trypsin
  • Trypsin Inhibitors
  • Journal Article

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