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

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.
Author Magdalena Filipowicz PChB
Magdalena Filipowicz,,
- Laboratory of Bioorganic Chemistry
, Natalia Ptaszyńska PChB
Natalia Ptaszyńska,,
- Laboratory of Bioorganic Chemistry
, Katarzyna Olkiewicz PChB
Katarzyna Olkiewicz ,,
- Laboratory of Bioorganic Chemistry
, Dawid Dębowski PChB
Dawid Dębowski,,
- Laboratory of Bioorganic Chemistry
, Kamila Ćwikłowska
Kamila Ćwikłowska,,
-
, Timo Burster
Timo Burster,,
-
, Michał Pikuła
Michał Pikuła,,
-
, Adam Krzystyniak
Adam Krzystyniak,,
-
, Anna Łęgowska PChB
Anna Łęgowska,,
- Laboratory of Bioorganic Chemistry
, Krzysztof Rolka PChB
Krzysztof Rolka,,
- Laboratory of Bioorganic Chemistry
Journal seriesBiopolymers, ISSN 0006-3525
Issue year2017
Vol108
No2
Pages1-26
Publication size in sheets1.25
Keywords in Englishpeptide splicing, SFTI-1 analogues, inhibitors, proteolysis, FRET, RGD peptides
DOIDOI:10.1002/bip.22988
URL http://onlinelibrary.wiley.com/doi/10.1002/bip.22988/epdf
Languageen angielski
Score (nominal)25
ScoreMinisterial score = 25.0, 20-12-2017, ArticleFromJournal
Ministerial score (2013-2016) = 25.0, 20-12-2017, ArticleFromJournal
Publication indicators WoS Impact Factor: 2016 = 1.908 (2) - 2016=2.225 (5)
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