P209-M Non-Isobaric Triplex-Labeling Strategies for Proteomics

A non-isobaric label chemistry is introduced that allows the triplexed quantification of proteomic samples based on the isotope-coded protein-labeling technology (ICPL). Amino groups of intact proteins were derivatized with three isotopically different nicotinoyl reagents (L: 12C6H4; M: 12C6D4; and...

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Main Authors: Schweiger-Hufnagel, U., Behrens, M., Hahner, S., Keidel, E., Dosch, D., Martin, S., Lottspeich, F., Halder, T., Suckau, D., Baessmann, C.
Formato: Artigo
Idioma:English
Publicado em: The Association of Biomolecular Resource Facilities 2007
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Poster Abstracts: Quantitative Proteomics
Acesso em linha:https://ncbi.nlm.nih.gov/pmc/articles/PMC2291819/
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spelling pubmed-22918192008-05-27 P209-M Non-Isobaric Triplex-Labeling Strategies for Proteomics Schweiger-Hufnagel, U. Behrens, M. Hahner, S. Keidel, E. Dosch, D. Martin, S. Lottspeich, F. Halder, T. Suckau, D. Baessmann, C. J Biomol Tech Poster Abstracts: Quantitative Proteomics A non-isobaric label chemistry is introduced that allows the triplexed quantification of proteomic samples based on the isotope-coded protein-labeling technology (ICPL). Amino groups of intact proteins were derivatized with three isotopically different nicotinoyl reagents (L: 12C6H4; M: 12C6D4; and H: 13C6H4) prior to protein separation and digestion. Here, we describe different approaches in respect of MS instruments and protein separation. Protein samples with different complexity were labeled with ICPL. For one sample, the proteins were digested, separated by CAP LC, and spotted on disposable targets for subsequent MALDI measurements. The other sample was separated on a 1D gel, excised, digested, and supplied to LC-MS/MS using a Qq orthogonal time-of-flight instrument. Excellent results were obtained from the LC-MALDI and the Qq TOF approach with the three protein samples labeled differentially and measured together. The multiplexed labeling allows us to reduce the experimental effort when proteomics experiments are run as replicates and when more than two different states are to be compared. A unique property of ICPL compared to other label chemistries is its compatibility with protein pre-fractionation, as the labeling reaction works on the undigested protein. The Association of Biomolecular Resource Facilities 2007-02 /pmc/articles/PMC2291819/ Text en Copyright © Copyright 2007 by The Association of Biomolecular Resource Facilities
institution US National Library of Medicine
collection PubMed Central
language English
format Article
topic Poster Abstracts: Quantitative Proteomics
spellingShingle Poster Abstracts: Quantitative Proteomics
Schweiger-Hufnagel, U.
Behrens, M.
Hahner, S.
Keidel, E.
Dosch, D.
Martin, S.
Lottspeich, F.
Halder, T.
Suckau, D.
Baessmann, C.
P209-M Non-Isobaric Triplex-Labeling Strategies for Proteomics
description A non-isobaric label chemistry is introduced that allows the triplexed quantification of proteomic samples based on the isotope-coded protein-labeling technology (ICPL). Amino groups of intact proteins were derivatized with three isotopically different nicotinoyl reagents (L: 12C6H4; M: 12C6D4; and H: 13C6H4) prior to protein separation and digestion. Here, we describe different approaches in respect of MS instruments and protein separation. Protein samples with different complexity were labeled with ICPL. For one sample, the proteins were digested, separated by CAP LC, and spotted on disposable targets for subsequent MALDI measurements. The other sample was separated on a 1D gel, excised, digested, and supplied to LC-MS/MS using a Qq orthogonal time-of-flight instrument. Excellent results were obtained from the LC-MALDI and the Qq TOF approach with the three protein samples labeled differentially and measured together. The multiplexed labeling allows us to reduce the experimental effort when proteomics experiments are run as replicates and when more than two different states are to be compared. A unique property of ICPL compared to other label chemistries is its compatibility with protein pre-fractionation, as the labeling reaction works on the undigested protein.
author Schweiger-Hufnagel, U.
Behrens, M.
Hahner, S.
Keidel, E.
Dosch, D.
Martin, S.
Lottspeich, F.
Halder, T.
Suckau, D.
Baessmann, C.
author_facet Schweiger-Hufnagel, U.
Behrens, M.
Hahner, S.
Keidel, E.
Dosch, D.
Martin, S.
Lottspeich, F.
Halder, T.
Suckau, D.
Baessmann, C.
author_sort Schweiger-Hufnagel, U.
title P209-M Non-Isobaric Triplex-Labeling Strategies for Proteomics
title_short P209-M Non-Isobaric Triplex-Labeling Strategies for Proteomics
title_full P209-M Non-Isobaric Triplex-Labeling Strategies for Proteomics
title_fullStr P209-M Non-Isobaric Triplex-Labeling Strategies for Proteomics
title_full_unstemmed P209-M Non-Isobaric Triplex-Labeling Strategies for Proteomics
title_sort p209-m non-isobaric triplex-labeling strategies for proteomics
publisher The Association of Biomolecular Resource Facilities
publisher_facet The Association of Biomolecular Resource Facilities
publishDate 2007
url https://ncbi.nlm.nih.gov/pmc/articles/PMC2291819/
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