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
Publisher Wiley
Publication Place Weinheim
Publication Date 2015-01-26
Keyword(s) Droplet microfluidics
Mass spectrometry
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
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NEBIS System Number 000033716
Source Database ID SCOPUS-84921532199
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  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