Phospho-Tyrosine(s) vs. Phosphatidylinositol Binding in Shc Mediated Integrin Signaling


The Shc adaptor protein, particularly its p52 isoform, has been identified as a primary signaling partner for the tyrosine(s)-phosphorylated cytoplasmic tails of activated β3 integrins. Inspired by our recent structure of the Shc PTB domain in complex with a bi-phosphorylated peptide derived from β3 cytoplasmic tail, we have initiated the investigation of Shc interaction with phospholipids of the membrane. We are particularly focused on PtdIns and their effects on Shc mediated integrin signaling in vitro. Here we present thermodynamic profiles and molecular details of the interactions between Shc, integrin, and PtdIns, all of which have been studied by ITC and solution NMR methods. A model of p52 Shc interaction with phosphorylated β3 integrin cytoplasmic tail at the cytosolic face of the plasma membrane is proposed based on these data.

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Lin, X. and Vinogradova, O. (2015) Phospho-Tyrosine(s) vs. Phosphatidylinositol Binding in Shc Mediated Integrin Signaling. American Journal of Molecular Biology, 5, 17-31. doi: 10.4236/ajmb.2015.52003.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Vinogradova, O., et al. (2002) A Structural Mechanism of Integrin Alpha(IIb)Beta(3) “Inside-Out” Activation as Regulated by Its Cytoplasmic Face. Cell, 110, 587-597.
[2] Vinogradova, O., et al. (2004) Membrane-Mediated Structural Transitions at the Cytoplasmic Face during Integrin Activation. Proceedings of the National Academy of Sciences USA, 101, 4094-4099.
[3] Deshmukh, L., et al. (2011) Tyrosine Phosphorylation as a Conformational Switch: A Case Study of Integrin Beta3 Cytoplasmic Tail. Journal of Biological Chemistry, 286, 40943-40953.
[4] Deshmukh, L., Gorbatyuk, V. and Vinogradova, O. (2010) Integrin Beta3 Phosphorylation Dictates Its Complex with Shc PTB Domain. Journal of Biological Chemistry, 285, 34875-24884.
[5] Schaffner-Reckinger, E., et al. (1998) Distinct Involvement of Beta3 Integrin Cytoplasmic Domain Tyrosine Residues 747 and 759 in Integrin-Mediated Cytoskeletal Assembly and Phosphotyrosine Signaling. Journal of Biological Chemistry, 273, 12623-12632.
[6] Jenkins, A.L., et al. (1998) Tyrosine Phosphorylation of the Beta3 Cytoplasmic Domain Mediates Integrin-Cytoskeletal Interactions. Journal of Biological Chemistry, 273, 13878-13885.
[7] Phillips, D.R., et al. (2001) Integrin Tyrosine Phosphorylation in Platelet Signaling. Current Opinion in Cell Biology, 13, 546-554.
[8] Law, D.A., et al. (1999) Integrin Cytoplasmic Tyrosine Motif Is Required for Outside-In AlphaIIbbeta3 Signalling and Platelet Function. Nature, 401, 808-811.
[9] Cowan, K.J., Law, D.A. and Phillips, D.R. (2000) Identification of Shc as the Primary Protein Binding to the Tyrosine-Phosphorylated Beta3 Subunit of Alpha IIbbeta 3 during Outside-In Integrin Platelet Signaling. Journal of Biological Chemistry, 275, 36423-36429.
[10] Higashi, T., et al. (2004) Direct Demonstration of Involvement of the Adaptor Protein ShcA in the Regulation of Ca2+-Induced Platelet Aggregation. Biochemical and Biophysical Research Communications, 322, 700-704.
[11] Pelicci, G., Lanfrancone, L., Grignani, F., McGlade, J., Cavallo, F., Forni, G., et al. (1992) A Novel Transforming Protein (SHC) with an SH2 Domain Is Implicated in Mitogenic Signal Transduction. Cell, 70, 93-104.
[12] Ravichandran, K.S. (2001) Signaling via Shc Family Adapter Proteins. Oncogene, 20, 6322-6330.
[13] Rameh, L.E., Chen, C.S. and Cantley, L.C. (1995) Phosphatidylinositol (3, 4, 5)P3 Interacts with SH2 Domains and Modulates PI 3-Kinase Association with Tyrosine-Phosphorylated Proteins. Cell, 83, 821-830.
[14] Balla, T. (2005) Inositol-Lipid Binding Motifs: Signal Integrators through Protein-Lipid and Protein-Protein Interactions. Journal of Cell Science, 118, 2093-2104.
[15] Zhou, M.M., Ravichandran, K.S., Olejniczak, E.T., Petros, A.M., Meadows, R.P., Sattler, M., et al. (1995) Structure and Ligand Recognition of the Phosphotyrosine Binding Domain of Shc. Nature, 378, 584-592.
[16] DiNitto, J.P. and Lambright, D.G. (2006) Membrane and Juxtamembrane Targeting by PH and PTB Domains. Biochimica et Biophysica Acta, 1761, 850-867.
[17] Farooq, A. and Zhou, M.M. (2004) PTB or Not to Be: Promiscuous, Tolerant and Bizarro Domains Come of Age. IUBMB Life, 56, 547-557.
[18] Farooq, A., Plotnikova, O., Zeng, L. and Zhou, M.-M. (1999) Phosphotyrosine Binding Domains of Shc and Insulin Receptor Substrate 1 Recognize the NPXpY Motif in a Thermodynamically Distinct Manner. The Journal of Biological Chemistry, 274, 6114-6121.
[19] Zhou, M.M., Harlan, J.E., Wade, W.S., Crosby, S., Ravichandran, K.S., Burakoff, S.J. and Fesik, S.W. (1995) Binding Affinities of Tyrosine-Phosphorylated Peptides to the COOH-Terminal SH2 and NH2-Terminal Phosphotyrosine Binding Domains of Shc. The Journal of Biological Chemistry, 270, 31119-31123.
[20] Ravichandran, K.S., Zhou, M.M., Pratt, J.C., Harlan, J.E., Walk, S.F., Fesik, S.W. and Burakoff, S.J. (1997) Evidence for a Requirement for both Phospholipid and Phosphotyrosine Binding via the Shc Phosphotyrosine-Binding Domain in Vivo. Molecular and Cellular Biology, 17, 5540-5549.
[21] George, R., Schuller, A.C., Harris, R. and Ladbury, J.E. (2008) A Phosphorylation-Dependent Gating Mechanism Controls the SH2 Domain Interactions of the Shc Adaptor Protein. Journal of Molecular Biology, 377, 740-747.
[22] Delaglio, F., Grzesiek, S., Vuister, G.W., Zhu, G., Pfeifer, J. and Bax, A. (1995) NMRPipe: A Multidimensional Spectral Processing System Based on UNIX Pipes. Journal of Biomolecular NMR, 6, 277-293.
[23] Vranken, W.F., Boucher, W., Stevens, T.J., Fogh, R.H., Pajon, A., Llinas, M., et al. (2005) The CCPN Data Model for NMR Spectroscopy: Development of a Software Pipeline. Proteins, 59, 687-696.
[24] Vinogradova, O. and Qin, J. (2012) NMR as a Unique Tool in Assessment and Complex Determination of Weak Protein-Protein Interactions. Topics in Current Chemistry, 326, 35-45.
[25] Perozzo, R., Folkers, G. and Scapozza, L. (2004) Thermodynamics of Protein-Ligand Interactions: History, Presence, and Future Aspects. Journal of Receptors and Signal Transduction, 24, 1-52.
[26] Katyal, P., Puthenveetil, R. and Vinogradova, O. (2013) Structural Insights into the Recognition of β3 Integrin Cytoplasmic Tail by SH3 Domain of Src Kinase. Protein Science, 22, 1358-1365.
[27] Ursini-Siegel, J., Hardy, W.R., Zuo, D., Lam, S.H., Sanguin-Gendreau, V., Cardiff, R.D., et al. (2008) ShcA Signalling Is Essential for Tumour Progression in Mouse Models of Human Breast Cancer. EMBO Journal, 27, 910-920.
[28] Pettersen, E.F., Goddard, T.D., Huang, C.C., Couch, G.S., Greenblatt, D.M., Meng, E.C. and Ferrin, T.E. (2004) UCSF Chimera—A Visualization System for Exploratory Research and Analysis. Journal of Computational Chemistry, 25, 1605-1612.
[29] Gurtovenko, A.A. and Vattulainen, I. (2007) Lipid Transmembrane Asymmetry and Intrinsic Membrane Potential: Two Sides of the Same Coin. Journal of the American Chemical Society, 129, 5358-5359.

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