This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Supplemental material
Right arrow An erratum has been published
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Smith, M. J.
Right arrow Articles by Pawson, T.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Smith, M. J.
Right arrow Articles by Pawson, T.

 Previous Article  |  Next Article 

Molecular and Cellular Biology, November 2006, p. 8461-8474, Vol. 26, No. 22
0270-7306/06/$08.00+0     doi:10.1128/MCB.01491-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Screening for PTB Domain Binding Partners and Ligand Specificity Using Proteome-Derived NPXY Peptide Arrays{triangledown} ,{dagger}

Matthew J. Smith,1,2 W. Rod Hardy,1,2 James M. Murphy,1 Nina Jones,1 and Tony Pawson1,2*

Samuel Lunenfeld Research Institute, Mount Sinai Hospital, 600 University Avenue, Toronto, Ontario, Canada M5G 1X5,1 Department of Molecular and Medical Genetics, University of Toronto, Toronto, Canada2

Received 10 August 2006/ Returned for modification 21 August 2006/ Accepted 31 August 2006

Modular interaction domains that recognize peptide motifs in target proteins can impart selectivity in signaling pathways. Phosphotyrosine binding (PTB) domains are components of cytoplasmic docking proteins that bind cell surface receptors through NPXY motifs. We have employed a library of human proteome-derived NXXY sequences to explore PTB domain specificity and function. SPOTS peptide arrays were used to create a comprehensive matrix of receptor motifs that were probed with a set of 10 diverse PTB domains. This approach confirmed that individual PTB domains have selective and distinct recognition properties and provided a means to explore over 2,500 potential PTB domain-NXXY interactions. The results correlated well with previously known associations between full-length proteins and predicted novel interactions, as well as consensus binding data for specific PTB domains. Using the Ret, MuSK, and ErbB2 receptor tyrosine kinases, we show that interactions of these receptors with PTB domains predicted to bind by the NXXY arrays do occur in cells. Proteome-based peptide arrays can therefore identify networks of receptor interactions with scaffold proteins that may be physiologically relevant.

* Corresponding author. Mailing address: Samuel Lunenfeld Research Institute, Mount Sinai Hospital, 600 University Avenue, Toronto, Ontario, Canada M5G 1X5. Phone: (416) 586-8262. Fax: (416) 586-8869. E-mail: pawson{at}mshri.on.ca.

{triangledown} Published ahead of print on 18 September 2006.

{dagger} Supplemental data for this article may be found at http://mcb.asm.org/.

Molecular and Cellular Biology, November 2006, p. 8461-8474, Vol. 26, No. 22
0270-7306/06/$08.00+0     doi:10.1128/MCB.01491-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Crose, L. E. S., Hilder, T. L., Sciaky, N., Johnson, G. L. (2009). Cerebral Cavernous Malformation 2 Protein Promotes Smad Ubiquitin Regulatory Factor 1-mediated RhoA Degradation in Endothelial Cells. J. Biol. Chem. 284: 13301-13305 [Abstract] [Full Text]  
  • Hanke, S., Mann, M. (2009). The Phosphotyrosine Interactome of the Insulin Receptor Family and Its Substrates IRS-1 and IRS-2. Mol. Cell. Proteomics 8: 519-534 [Abstract] [Full Text]  
  • Miller, M. L., Jensen, L. J., Diella, F., Jorgensen, C., Tinti, M., Li, L., Hsiung, M., Parker, S. A., Bordeaux, J., Sicheritz-Ponten, T., Olhovsky, M., Pasculescu, A., Alexander, J., Knapp, S., Blom, N., Bork, P., Li, S., Cesareni, G., Pawson, T., Turk, B. E., Yaffe, M. B., Brunak, S., Linding, R. (2008). Linear Motif Atlas for Phosphorylation-Dependent Signaling. Sci Signal 1: ra2-ra2 [Abstract] [Full Text]  
  • Radzimanowski, J., Ravaud, S., Schlesinger, S., Koch, J., Beyreuther, K., Sinning, I., Wild, K. (2008). Crystal Structure of the Human Fe65-PTB1 Domain. J. Biol. Chem. 283: 23113-23120 [Abstract] [Full Text]  
  • Li, L., Wu, C., Huang, H., Zhang, K., Gan, J., Li, S. S.-C. (2008). Prediction of phosphotyrosine signaling networks using a scoring matrix-assisted ligand identification approach. Nucleic Acids Res 36: 3263-3273 [Abstract] [Full Text]  
  • Northey, J. J., Chmielecki, J., Ngan, E., Russo, C., Annis, M. G., Muller, W. J., Siegel, P. M. (2008). Signaling through ShcA Is Required for Transforming Growth Factor {beta}- and Neu/ErbB-2-Induced Breast Cancer Cell Motility and Invasion. Mol. Cell. Biol. 28: 3162-3176 [Abstract] [Full Text]  
  • Mishra, S. K., Jha, A., Steinhauser, A. L., Kokoza, V. A., Washabaugh, C. H., Raikhel, A. S., Foster, W. A., Traub, L. M. (2008). Internalization of LDL-receptor superfamily yolk-protein receptors during mosquito oogenesis involves transcriptional regulation of PTB-domain adaptors. J. Cell Sci. 121: 1264-1274 [Abstract] [Full Text]  
  • Jones, N., Hardy, W. R., Friese, M. B., Jorgensen, C., Smith, M. J., Woody, N. M., Burden, S. J., Pawson, T. (2007). Analysis of a Shc Family Adaptor Protein, ShcD/Shc4, That Associates with Muscle-Specific Kinase. Mol. Cell. Biol. 27: 4759-4773 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»