Chemical derivatization of phosphoserine and phosphothreonine containing peptides to increase sensitivity for MALDI-based analysis and for selectivity of MS/MS analysis
Arrigoni, Giorgio LU ; Resjö, Svante LU ; Levander, Fredrik LU ; Nilsson, R ; Degerman, Eva LU
; Quadroni, M
; Pinna, LA
and James, Peter
LU
(2006)
In Proteomics
6(3).
p.757-766
- Abstract
- Protein phosphorylation is one of the most important and common ways of regulating protein function in cells. However, phosphopeptides are difficult to analyse, ionising poorly under standard MALDI conditions. Several methods have been developed to deal with the low sensitivity and specificity of phosphopeptide analysis. Here, we show an approach using a simple one-step beta-elimination/Michael addition reaction for the derivatization of phosphoserine and phosphothreonine. The substitution of the negatively charged phosphate group by a positively charged S-ethylpyridyl group greatly improves the ionisation of the modified peptides, especially in MALDI MS, increasing the sensitivity of the analysis. The modification allows the formation of... (More)
- Protein phosphorylation is one of the most important and common ways of regulating protein function in cells. However, phosphopeptides are difficult to analyse, ionising poorly under standard MALDI conditions. Several methods have been developed to deal with the low sensitivity and specificity of phosphopeptide analysis. Here, we show an approach using a simple one-step beta-elimination/Michael addition reaction for the derivatization of phosphoserine and phosphothreonine. The substitution of the negatively charged phosphate group by a positively charged S-ethylpyridyl group greatly improves the ionisation of the modified peptides, especially in MALDI MS, increasing the sensitivity of the analysis. The modification allows the formation of a unique fragment ion at m/z 106 under mild collisional activation conditions, which can be used for parent (precursor) ion scanning in order to improve both the sensitivity and the selectivity of the analysis. The optimisation of the approach is described for a standard model peptide and protein and then applied to phosphorylation analysis in two biologically derived proteins purified from different experimental systems. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/417345
- author
- Arrigoni, Giorgio
LU
; Resjö, Svante
LU
; Levander, Fredrik
LU
; Nilsson, R
; Degerman, Eva
LU
; Quadroni, M
; Pinna, LA
and James, Peter
LU
- organization
- publishing date
- 2006
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- matrix-assisted laser desorption/ionization time of, derivatization, flight mass spectrometry, phosphorylation, sensitivity, spectrometry, tandem mass
- in
- Proteomics
- volume
- 6
- issue
- 3
- pages
- 757 - 766
- publisher
- John Wiley & Sons Inc.
- external identifiers
-
- pmid:16372258
- wos:000235414600003
- scopus:32944464615
- ISSN
- 1615-9861
- DOI
- 10.1002/pmic.200500073
- language
- English
- LU publication?
- yes
- id
- 921ec828-a57b-4bf5-abc1-f8f519bda4f5 (old id 417345)
- date added to LUP
- 2016-04-01 12:21:28
- date last changed
- 2023-09-02 04:49:44
@article{921ec828-a57b-4bf5-abc1-f8f519bda4f5,
abstract = {{Protein phosphorylation is one of the most important and common ways of regulating protein function in cells. However, phosphopeptides are difficult to analyse, ionising poorly under standard MALDI conditions. Several methods have been developed to deal with the low sensitivity and specificity of phosphopeptide analysis. Here, we show an approach using a simple one-step beta-elimination/Michael addition reaction for the derivatization of phosphoserine and phosphothreonine. The substitution of the negatively charged phosphate group by a positively charged S-ethylpyridyl group greatly improves the ionisation of the modified peptides, especially in MALDI MS, increasing the sensitivity of the analysis. The modification allows the formation of a unique fragment ion at m/z 106 under mild collisional activation conditions, which can be used for parent (precursor) ion scanning in order to improve both the sensitivity and the selectivity of the analysis. The optimisation of the approach is described for a standard model peptide and protein and then applied to phosphorylation analysis in two biologically derived proteins purified from different experimental systems.}},
author = {{Arrigoni, Giorgio and Resjö, Svante and Levander, Fredrik and Nilsson, R and Degerman, Eva and Quadroni, M and Pinna, LA and James, Peter}},
issn = {{1615-9861}},
keywords = {{matrix-assisted laser desorption/ionization time of; derivatization; flight mass spectrometry; phosphorylation; sensitivity; spectrometry; tandem mass}},
language = {{eng}},
number = {{3}},
pages = {{757--766}},
publisher = {{John Wiley & Sons Inc.}},
series = {{Proteomics}},
title = {{Chemical derivatization of phosphoserine and phosphothreonine containing peptides to increase sensitivity for MALDI-based analysis and for selectivity of MS/MS analysis}},
url = {{http://dx.doi.org/10.1002/pmic.200500073}},
doi = {{10.1002/pmic.200500073}},
volume = {{6}},
year = {{2006}},
}