PMID- 23783835
OWN - NLM
STAT- MEDLINE
DCOM- 20140529
LR  - 20240330
IS  - 1618-2650 (Electronic)
IS  - 1618-2642 (Print)
IS  - 1618-2642 (Linking)
VI  - 405
IP  - 28
DP  - 2013 Nov
TI  - Synthesis and characterisation of PEG-peptide surfaces for proteolytic enzyme 
      detection.
PG  - 9049-59
LID - 10.1007/s00216-013-7082-z [doi]
AB  - 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.
FAU - Trzcinska, Roza
AU  - Trzcinska R
AD  - Centre of Polymer and Carbon Materials, Polish Academy of Sciences, M. 
      Curie-Sklodowskiej 34, Zabrze, 41-819, Poland.
FAU - Suder, Piotr
AU  - Suder P
FAU - Bodzon-Kulakowska, Anna
AU  - Bodzon-Kulakowska A
FAU - Skalska, Magdalena
AU  - Skalska M
FAU - Marcinkowski, Andrzej
AU  - Marcinkowski A
FAU - Kubacki, Jerzy
AU  - Kubacki J
FAU - Pedrys, Roman
AU  - Pedrys R
FAU - Silberring, Jerzy
AU  - Silberring J
FAU - Dworak, Andrzej
AU  - Dworak A
FAU - Trzebicka, Barbara
AU  - Trzebicka B
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20130620
PL  - Germany
TA  - Anal Bioanal Chem
JT  - Analytical and bioanalytical chemistry
JID - 101134327
RN  - 0 (Peptides)
RN  - 3WJQ0SDW1A (Polyethylene Glycols)
RN  - EC 3.4.- (Peptide Hydrolases)
SB  - IM
MH  - Animals
MH  - Biosensing Techniques/instrumentation/*methods
MH  - Humans
MH  - Peptide Hydrolases/*analysis
MH  - Peptides/chemical synthesis/*chemistry
MH  - Polyethylene Glycols/*chemistry
PMC - PMC3825591
EDAT- 2013/06/21 06:00
MHDA- 2014/05/30 06:00
PMCR- 2013/06/20
CRDT- 2013/06/21 06:00
PHST- 2013/03/20 00:00 [received]
PHST- 2013/05/17 00:00 [accepted]
PHST- 2013/05/10 00:00 [revised]
PHST- 2013/06/21 06:00 [entrez]
PHST- 2013/06/21 06:00 [pubmed]
PHST- 2014/05/30 06:00 [medline]
PHST- 2013/06/20 00:00 [pmc-release]
AID - 7082 [pii]
AID - 10.1007/s00216-013-7082-z [doi]
PST - ppublish
SO  - Anal Bioanal Chem. 2013 Nov;405(28):9049-59. doi: 10.1007/s00216-013-7082-z. Epub 
      2013 Jun 20.