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Synthesis and characterisation of PEG-peptide surfaces for proteolytic enzyme detection


Synthesis and characterisation of PEG-peptide surfaces for proteolytic enzyme detection

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dc.contributor.author Trzcinska, Roza pl
dc.contributor.author Suder, Piotr pl
dc.contributor.author Bodzon-Kulakowska, Anna pl
dc.contributor.author Skalska, Magdalena pl
dc.contributor.author Marcinkowski, Andrzej pl
dc.contributor.author Kubacki, Jerzy pl
dc.contributor.author Pędrys, Roman [SAP11006083] pl
dc.contributor.author Silberring, Jerzy [SAP11016264] pl
dc.contributor.author Dworak, Andrzej pl
dc.contributor.author Trzebicka, Barbara pl
dc.date.accessioned 2015-06-29T18:25:57Z
dc.date.available 2015-06-29T18:25:57Z
dc.date.issued 2013 pl
dc.identifier.issn 1618-2642 pl
dc.identifier.uri http://ruj.uj.edu.pl/xmlui/handle/item/10711
dc.language eng pl
dc.rights Udzielam licencji. Uznanie autorstwa *
dc.rights.uri http://creativecommons.org/licenses *
dc.title Synthesis and characterisation of PEG-peptide surfaces for proteolytic enzyme detection pl
dc.type JournalArticle pl
dc.description.physical 9049-9059 pl
dc.abstract.en Peptide surfaces were obtained by the covalent immobilisation of fluorescently labelled pentapeptides carboxyfluorescein–glycine–arginine–methionine–leucine–glycine, either directly or through a poly(ethylene glycol) (PEG) linker on modified silicon wafers. Each step during the preparation of the peptide surfaces was confirmed by several surface characterisation techniques. Time-of-flight secondary ion mass spectrometry (ToF-SIMS) and X-ray photoelectron spectroscopy were used to determine the surface composition, the wafers philicity was measured by contact angle and atomic force microscopy was used to investigate the surface morphology. Exposure of the peptide surfaces to trypsin resulted in the release of a fluorescently labelled peptide product, which allowed the kinetics of the enzymatic reaction to be followed with the aid of fluorescence spectroscopy. The electrospray ionisation mass spectrometry analysis of the post-digestion solution confirmed that the pentapeptides attached to the solid support undergo specific trypsin hydrolysis at the C-terminus of the arginine residues. Detailed surface analyses before and after the enzyme action was performed using ToF-SIMS. Because of the limited accessibility of the short peptide directly attached to the surface, a quantitative yield of enzymatic hydrolysis was observed only in case when the peptide was bound through the PEG linker. The insertion of the PEG linker increased the number of immobilised peptides and the rate of enzymatic digestion which consequently improved the quality of the enzyme assays. The described approach may be used for different peptide sequences designed for other proteases. Figure Monitoring of trypsin hydrolysis on PEG-peptide surface pl
dc.subject.en amino acids/peptides pl
dc.subject.en biomaterials pl
dc.subject.en interface/surface analysis pl
dc.subject.en mass spectrometry/ICP-MS pl
dc.subject.en polymers pl
dc.description.volume 405 pl
dc.description.number 28 pl
dc.description.publication 1 pl
dc.identifier.doi 10.1007/s00216-013-7082-z pl
dc.identifier.eissn 1618-2650 pl
dc.title.journal Analytical and Bioanalytical Chemistry pl
dc.language.container eng pl
dc.affiliation Wydział Fizyki, Astronomii i Informatyki Stosowanej : Instytut Fizyki im. Mariana Smoluchowskiego pl
dc.subtype Article pl
dc.rights.original CC-BY; inne; ostateczna wersja wydawcy; w momencie opublikowania; 0; pl
dc.identifier.project ROD UJ / P pl
.pointsMNiSW [2013 A]: 40

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Udzielam licencji. Uznanie autorstwa Except where otherwise noted, this item's license is described as Udzielam licencji. Uznanie autorstwa