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

doi:10.1007/s00216-013-7082-z

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

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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