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A Cross-Species Study of PI3K Protein-Protein Interactions Reveals the Direct Interaction of P85 and SHP2
Using a series of immunoprecipitation (IP) – tandem mass spectrometry (LC-MS/MS) experiments and reciprocal BLAST, we conducted a fly-human cross-species comparison of the phosphoinositide-3-kinase (PI3K) interactome in a drosophila S2R+ cell line and several NSCLC and human multiple myeloma cell li...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4738311/ https://www.ncbi.nlm.nih.gov/pubmed/26839216 http://dx.doi.org/10.1038/srep20471 |
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author | Breitkopf, Susanne B. Yang, Xuemei Begley, Michael J. Kulkarni, Meghana Chiu, Yu-Hsin Turke, Alexa B. Lauriol, Jessica Yuan, Min Qi, Jie Engelman, Jeffrey A. Hong, Pengyu Kontaridis, Maria I. Cantley, Lewis C. Perrimon, Norbert Asara, John M. |
author_facet | Breitkopf, Susanne B. Yang, Xuemei Begley, Michael J. Kulkarni, Meghana Chiu, Yu-Hsin Turke, Alexa B. Lauriol, Jessica Yuan, Min Qi, Jie Engelman, Jeffrey A. Hong, Pengyu Kontaridis, Maria I. Cantley, Lewis C. Perrimon, Norbert Asara, John M. |
author_sort | Breitkopf, Susanne B. |
collection | PubMed |
description | Using a series of immunoprecipitation (IP) – tandem mass spectrometry (LC-MS/MS) experiments and reciprocal BLAST, we conducted a fly-human cross-species comparison of the phosphoinositide-3-kinase (PI3K) interactome in a drosophila S2R+ cell line and several NSCLC and human multiple myeloma cell lines to identify conserved interacting proteins to PI3K, a critical signaling regulator of the AKT pathway. Using H929 human cancer cells and drosophila S2R+ cells, our data revealed an unexpected direct binding of Corkscrew, the drosophila ortholog of the non-receptor protein tyrosine phosphatase type II (SHP2) to the Pi3k21B (p60) regulatory subunit of PI3K (p50/p85 human ortholog) but no association with Pi3k92e, the human ortholog of the p110 catalytic subunit. The p85-SHP2 association was validated in human cell lines, and formed a ternary regulatory complex with GRB2-associated-binding protein 2 (GAB2). Validation experiments with knockdown of GAB2 and Far-Western blots proved the direct interaction of SHP2 with p85, independent of adaptor proteins and transfected FLAG-p85 provided evidence that SHP2 binding on p85 occurred on the SH2 domains. A disruption of the SHP2-p85 complex took place after insulin/IGF1 stimulation or imatinib treatment, suggesting that the direct SHP2-p85 interaction was both independent of AKT activation and positively regulates the ERK signaling pathway. |
format | Online Article Text |
id | pubmed-4738311 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-47383112016-02-09 A Cross-Species Study of PI3K Protein-Protein Interactions Reveals the Direct Interaction of P85 and SHP2 Breitkopf, Susanne B. Yang, Xuemei Begley, Michael J. Kulkarni, Meghana Chiu, Yu-Hsin Turke, Alexa B. Lauriol, Jessica Yuan, Min Qi, Jie Engelman, Jeffrey A. Hong, Pengyu Kontaridis, Maria I. Cantley, Lewis C. Perrimon, Norbert Asara, John M. Sci Rep Article Using a series of immunoprecipitation (IP) – tandem mass spectrometry (LC-MS/MS) experiments and reciprocal BLAST, we conducted a fly-human cross-species comparison of the phosphoinositide-3-kinase (PI3K) interactome in a drosophila S2R+ cell line and several NSCLC and human multiple myeloma cell lines to identify conserved interacting proteins to PI3K, a critical signaling regulator of the AKT pathway. Using H929 human cancer cells and drosophila S2R+ cells, our data revealed an unexpected direct binding of Corkscrew, the drosophila ortholog of the non-receptor protein tyrosine phosphatase type II (SHP2) to the Pi3k21B (p60) regulatory subunit of PI3K (p50/p85 human ortholog) but no association with Pi3k92e, the human ortholog of the p110 catalytic subunit. The p85-SHP2 association was validated in human cell lines, and formed a ternary regulatory complex with GRB2-associated-binding protein 2 (GAB2). Validation experiments with knockdown of GAB2 and Far-Western blots proved the direct interaction of SHP2 with p85, independent of adaptor proteins and transfected FLAG-p85 provided evidence that SHP2 binding on p85 occurred on the SH2 domains. A disruption of the SHP2-p85 complex took place after insulin/IGF1 stimulation or imatinib treatment, suggesting that the direct SHP2-p85 interaction was both independent of AKT activation and positively regulates the ERK signaling pathway. Nature Publishing Group 2016-02-03 /pmc/articles/PMC4738311/ /pubmed/26839216 http://dx.doi.org/10.1038/srep20471 Text en Copyright © 2016, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Breitkopf, Susanne B. Yang, Xuemei Begley, Michael J. Kulkarni, Meghana Chiu, Yu-Hsin Turke, Alexa B. Lauriol, Jessica Yuan, Min Qi, Jie Engelman, Jeffrey A. Hong, Pengyu Kontaridis, Maria I. Cantley, Lewis C. Perrimon, Norbert Asara, John M. A Cross-Species Study of PI3K Protein-Protein Interactions Reveals the Direct Interaction of P85 and SHP2 |
title | A Cross-Species Study of PI3K Protein-Protein Interactions Reveals the Direct Interaction of P85 and SHP2 |
title_full | A Cross-Species Study of PI3K Protein-Protein Interactions Reveals the Direct Interaction of P85 and SHP2 |
title_fullStr | A Cross-Species Study of PI3K Protein-Protein Interactions Reveals the Direct Interaction of P85 and SHP2 |
title_full_unstemmed | A Cross-Species Study of PI3K Protein-Protein Interactions Reveals the Direct Interaction of P85 and SHP2 |
title_short | A Cross-Species Study of PI3K Protein-Protein Interactions Reveals the Direct Interaction of P85 and SHP2 |
title_sort | cross-species study of pi3k protein-protein interactions reveals the direct interaction of p85 and shp2 |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4738311/ https://www.ncbi.nlm.nih.gov/pubmed/26839216 http://dx.doi.org/10.1038/srep20471 |
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