Characterization of the phosphotyrosine-binding domain of the Drosophila Shc protein.

The Journal of Biological Chemistry
S C LiT Pawson

Abstract

The phosphotyrosine-binding (PTB) domain of Drosophila Shc (dShc) binds in vitro to phosphopeptides containing the sequence motif NPXpY, and physically associates with the activated Drosophila epidermal growth factor receptor homologue (DER) in vivo. The structural elements, specificity and binding kinetics of the dShc PTB domain have now been characterized. The dShc PTB domain appeared similar to the insulin-like receptor substrate-1 PTB domain in secondary structure as suggested by Fourier transform infrared spectroscopy. Surface plasmon resonance measurements indicated that the dShc PTB domain bound with high affinity to phosphopeptides (Der) derived from the Tyr1228 site of the DER receptor. The kinetics of the dShc PTB domain-Der phosphopeptide interaction differed from those of a typical SH2 domain-ligand interaction, in that the PTB domain displayed slower on/off rates. Competition binding assays using truncated versions of the Der peptides revealed that high affinity binding to the dShc PTB domain requires, in addition to the NPXpY motif, the presence of hydrophobic residues at both positions -5 and -7 relative to phosphotyrosine. The dShc PTB domain showed a similar binding specificity to the human Shc (hShc) PTB domai...Continue Reading

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Citations

Jul 8, 1997·Proceedings of the National Academy of Sciences of the United States of America·S C LiT Pawson
Jun 18, 1999·Cellular Signalling·L WunderlichL Buday
Dec 29, 2004·Mechanisms of Ageing and Development·Orli SagiVadim Fraifeld
Jan 25, 2011·Analytical Biochemistry·Jun YokoyamaTakanori Kigawa
Jul 15, 2009·Biomaterials·Golnaz KaroubiDavid W Courtman
Dec 9, 2017·Molecular & Cellular Proteomics : MCP·Ran WeiShawn S-C Li
May 8, 2002·Nature Reviews. Molecular Cell Biology·Michael B Yaffe
Dec 13, 2005·The Journal of Immunology : Official Journal of the American Association of Immunologists·Sigal GelkopNoah Isakov
Nov 1, 2018·Molecular & Cellular Proteomics : MCP·Ran WeiShawn S-C Li

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