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Comparison of Three Quantitative Phosphoproteomic Strategies to Study Receptor Tyrosine Kinase Signaling
[Image: see text] There are three quantitative phosphoproteomic strategies most commonly used to study receptor tyrosine kinase (RTK) signaling. These strategies quantify changes in: (1) all three forms of phosphosites (phosphoserine, phosphothreonine and phosphotyrosine) following enrichment of pho...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical Society
2011
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3229182/ https://www.ncbi.nlm.nih.gov/pubmed/22013880 http://dx.doi.org/10.1021/pr200697x |
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author | Zhang, Guoan Neubert, Thomas A. |
author_facet | Zhang, Guoan Neubert, Thomas A. |
author_sort | Zhang, Guoan |
collection | PubMed |
description | [Image: see text] There are three quantitative phosphoproteomic strategies most commonly used to study receptor tyrosine kinase (RTK) signaling. These strategies quantify changes in: (1) all three forms of phosphosites (phosphoserine, phosphothreonine and phosphotyrosine) following enrichment of phosphopeptides by titanium dioxide or immobilized metal affinity chromatography; (2) phosphotyrosine sites following anti- phosphotyrosine antibody enrichment of phosphotyrosine peptides; or (3) phosphotyrosine proteins and their binding partners following anti-phosphotyrosine protein immunoprecipitation. However, it is not clear from literature which strategy is more effective. In this study, we assessed the utility of these three phosphoproteomic strategies in RTK signaling studies by using EphB receptor signaling as an example. We used all three strategies with stable isotope labeling with amino acids in cell culture (SILAC) to compare changes in phosphoproteomes upon EphB receptor activation. We used bioinformatic analysis to compare results from the three analyses. Our results show that the three strategies provide complementary information about RTK pathways. |
format | Online Article Text |
id | pubmed-3229182 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | American Chemical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-32291822011-12-05 Comparison of Three Quantitative Phosphoproteomic Strategies to Study Receptor Tyrosine Kinase Signaling Zhang, Guoan Neubert, Thomas A. J Proteome Res [Image: see text] There are three quantitative phosphoproteomic strategies most commonly used to study receptor tyrosine kinase (RTK) signaling. These strategies quantify changes in: (1) all three forms of phosphosites (phosphoserine, phosphothreonine and phosphotyrosine) following enrichment of phosphopeptides by titanium dioxide or immobilized metal affinity chromatography; (2) phosphotyrosine sites following anti- phosphotyrosine antibody enrichment of phosphotyrosine peptides; or (3) phosphotyrosine proteins and their binding partners following anti-phosphotyrosine protein immunoprecipitation. However, it is not clear from literature which strategy is more effective. In this study, we assessed the utility of these three phosphoproteomic strategies in RTK signaling studies by using EphB receptor signaling as an example. We used all three strategies with stable isotope labeling with amino acids in cell culture (SILAC) to compare changes in phosphoproteomes upon EphB receptor activation. We used bioinformatic analysis to compare results from the three analyses. Our results show that the three strategies provide complementary information about RTK pathways. American Chemical Society 2011-10-20 2011-12-02 /pmc/articles/PMC3229182/ /pubmed/22013880 http://dx.doi.org/10.1021/pr200697x Text en Copyright © 2011 American Chemical Society http://pubs.acs.org This is an open-access article distributed under the ACS AuthorChoice Terms & Conditions. Any use of this article, must conform to the terms of that license which are available at http://pubs.acs.org. |
spellingShingle | Zhang, Guoan Neubert, Thomas A. Comparison of Three Quantitative Phosphoproteomic Strategies to Study Receptor Tyrosine Kinase Signaling |
title | Comparison of Three Quantitative Phosphoproteomic Strategies to Study Receptor Tyrosine Kinase Signaling |
title_full | Comparison of Three Quantitative Phosphoproteomic Strategies to Study Receptor Tyrosine Kinase Signaling |
title_fullStr | Comparison of Three Quantitative Phosphoproteomic Strategies to Study Receptor Tyrosine Kinase Signaling |
title_full_unstemmed | Comparison of Three Quantitative Phosphoproteomic Strategies to Study Receptor Tyrosine Kinase Signaling |
title_short | Comparison of Three Quantitative Phosphoproteomic Strategies to Study Receptor Tyrosine Kinase Signaling |
title_sort | comparison of three quantitative phosphoproteomic strategies to study receptor tyrosine kinase signaling |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3229182/ https://www.ncbi.nlm.nih.gov/pubmed/22013880 http://dx.doi.org/10.1021/pr200697x |
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