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 - [Universitätsklinikum Ulm]
Kamila Ćwikłowska,,
, Timo Burster - [Universitätsklinikum Ulm]
Timo Burster,,
, Michał Pikuła - [Gdanski Uniwersytet Medyczny]
Michał Pikuła,,
- Gdanski Uniwersytet Medyczny
, Adam Krzystyniak - [Nencki Institute of Experimental Biology of the Polish Academy of Sciences]
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
Article numbere22988
Keywords in Englishpeptide splicing, SFTI-1 analogues, inhibitors, proteolysis, FRET, RGD peptides
ASJC Classification2700 General Medicine; 1303 Biochemistry; 1304 Biophysics; 1605 Organic Chemistry; 2502 Biomaterials
URL http://onlinelibrary.wiley.com/doi/10.1002/bip.22988/epdf
Languageen angielski
Score (nominal)25
Score sourcejournalList
ScoreMinisterial score = 25.0, 03-04-2020, ArticleFromJournal
Publication indicators WoS Citations = 3; Scopus Citations = 4; Scopus SNIP (Source Normalised Impact per Paper): 2017 = 0.621; WoS Impact Factor: 2017 = 1.99 (2) - 2017=2.165 (5)
Citation count*4 (2020-04-05)
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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.
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