title: Spliced analogues of trypsin inhibitor SFTI-1 and their application for tracing proteolysis and delivery of cargos inside the cells.
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.
authors.affiliation: Faculty of Chemistry, University of Gdansk, Gdansk, Poland.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Filipowicz
authors.affiliation: Faculty of Chemistry, University of Gdansk, Gdansk, Poland.
authors.forename: Natalia
authors.initials: N
authors.lastname: Ptaszyńska
authors.affiliation: Faculty of Chemistry, University of Gdansk, Gdansk, Poland.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Olkiewicz
authors.affiliation: Faculty of Chemistry, University of Gdansk, Gdansk, Poland.
authors.forename: Dawid
authors.initials: D
authors.lastname: Dębowski
authors.affiliation: Department of Neurosurgery, Ulm University Medical Center, Ulm, Germany.
authors.forename: Kamila
authors.initials: K
authors.lastname: Ćwikłowska
authors.affiliation: Department of Neurosurgery, Ulm University Medical Center, Ulm, Germany.
authors.forename: Timo
authors.initials: T
authors.lastname: Burster
authors.affiliation: Department of Clinical Immunology and Transplantology, Medical University of Gdansk, Gdansk, Poland.
authors.forename: Michał
authors.initials: M
authors.lastname: Pikuła
authors.affiliation: Nencki Institute of Experimental Biology, Warsaw, Poland.
authors.forename: Adam
authors.initials: A
authors.lastname: Krzystyniak
authors.affiliation: Faculty of Chemistry, University of Gdansk, Gdansk, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Łęgowska
authors.affiliation: Faculty of Chemistry, University of Gdansk, Gdansk, Poland.
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Rolka
mesh_terms: D000595:Amino Acid Sequence; D002460:Cell Line; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism*; D013058:Mass Spectrometry; D008856:Microscopy, Fluorescence; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism; D010455:Peptides / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D059748:Proteolysis; D014357:Trypsin / Q000737:chemistry / Q000378:metabolism*; D014361:Trypsin Inhibitors / Q000737:chemistry / Q000378:metabolism*
chemical_list: D009842:Oligopeptides; D010455:Peptides; D014361:Trypsin Inhibitors; C047981:arginyl-glycyl-aspartic acid; D014357:Trypsin
keywords: FRET; RGD peptides; SFTI-1 analogues; inhibitors; peptide splicing; proteolysis