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


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.
Author Magdalena Filipowicz (FCh / DMB / LBCh)
Magdalena Filipowicz,,
- Laboratory of Bioorganic Chemistry
, Natalia Ptaszyńska (FCh / DMB / LBCh)
Natalia Ptaszyńska,,
- Laboratory of Bioorganic Chemistry
, Katarzyna Olkiewicz (FCh / DMB / LBCh)
Katarzyna Olkiewicz ,,
- Laboratory of Bioorganic Chemistry
, Dawid Dębowski (FCh / DMB / LBCh)
Dawid Dębowski,,
- Laboratory of Bioorganic Chemistry
, Kamila Ćwikłowska
Kamila Ćwikłowska,,
, Timo Burster
Timo Burster,,
, Michał Pikuła
Michał Pikuła,,
, Adam Krzystyniak
Adam Krzystyniak,,
, Anna Łęgowska (FCh / DMB / LBCh)
Anna Łęgowska,,
- Laboratory of Bioorganic Chemistry
, Krzysztof Rolka (FCh / DMB / LBCh)
Krzysztof Rolka,,
- Laboratory of Bioorganic Chemistry
Journal seriesBiopolymers, ISSN 0006-3525, e-ISSN 1097-0282, (A 25 pkt)
Issue year2017
Publication size in sheets1.25
Keywords in Englishpeptide splicing, SFTI-1 analogues, inhibitors, proteolysis, FRET, RGD peptides
ASJC Classification1605 Organic Chemistry; 2502 Biomaterials; 1303 Biochemistry; 2700 General Medicine; 1304 Biophysics
URL http://onlinelibrary.wiley.com/doi/10.1002/bip.22988/epdf
Languageen angielski
Score (nominal)25
ScoreMinisterial score = 25.0, 24-07-2019, ArticleFromJournal
Publication indicators WoS Citations = 3; Scopus SNIP (Source Normalised Impact per Paper): 2016 = 0.722; WoS Impact Factor: 2017 = 1.99 (2) - 2017=2.165 (5)
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* presented citation count is obtained through Internet information analysis and it is close to the number calculated by the Publish or Perish system.