Screening for protein phosphorylation using nanoscale reactions on microdroplet arrays

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Author(s) Küster, Simon K., Pabst, Martin, Zenobi, Renato, Dittrich, Petra S.
Publication Type Journal Items, Publication Status: Published
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Title Screening for protein phosphorylation using nanoscale reactions on microdroplet arrays
Author(s) Küster, Simon K.
Pabst, Martin
Zenobi, Renato
Dittrich, Petra S.
Journal or Series Title Angewandte Chemie - International Edition
Volume Number 54
Issue Number 5
Start Page 1671
End Page 1675
ISSN 1433-7851
0570-0833
1521-3773
Publisher Wiley
Publication Place Weinheim
Publication Date 2015-01-26
Keyword(s) Droplet microfluidics
Mass spectrometry
Microarrays
Protein modifications
Protein phosphorylation
Abstract We present a novel and straightforward screening method to detect protein phosphorylations in complex protein mixtures. A proteolytic digest is separated by a conventional nanoscale liquid chromatography (nano-LC) separation and the eluate is immediately compartmentalized into microdroplets, which are spotted on a microarray MALDI plate. Subsequently, the enzyme alkaline phosphatase is applied to every second microarray spot to remove the phosphate groups from phosphorylated peptides, which results in a mass shift of n×−80 Da. The MALDI-MS scan of the microarray is then evaluated by a software algorithm to automatically identify the phosphorylated peptides by exploiting the characteristic chromatographic peak profile induced by the phosphatase treatment. This screening method does not require extensive MS/MS experiments or peak list evaluation and can be easily extended to other enzymatic or chemical reactions.
DOI 10.1002/anie.201409440
Additional Notes Article first published online 12 December 2014
Document Type Article
Publication Status Published
Language English
Assigned Organisational Unit(s) 03807
03430
Organisational Unit(s)
NEBIS System Number 000033716
Source Database ID SCOPUS-84921532199
WOS-000348713900053
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@article{Kstr2015,
  author = "K{\"{u}}ster, Simon K. and Pabst, Martin and Zenobi, Renato and Dittrich, Petra S.",
  title = "{S}creening for protein phosphorylation using nanoscale reactions on microdroplet arrays",
  journal = "Angewandte Chemie - International Edition",
  year = 2015,
  volume = "54",
  number = "5",
  pages = "1671--1675",
  month = jan,
}


E-Citations record created: Tue, 13 Jan 2015, 10:17:51 CET