TY - JOUR
T1 - EGF Regulates the Interaction of Tks4 with Src through Its SH2 and SH3 Domains
AU - Dülk, Metta
AU - Szeder, Bálint
AU - Glatz, Gábor
AU - Merö, Balázs L.
AU - Koprivanacz, Kitti
AU - Kudlik, Gyöngyi
AU - Vas, Virág
AU - Sipeki, Szabolcs
AU - Cserkaszky, Anna
AU - Radnai, László
AU - Buday, László
N1 - Funding Information:
*E-mail: [email protected]. *E-mail: [email protected]. ORCID Laś zló Buday: 0000-0003-3518-5757 Present Address ⊥L.R.: Department of Molecular Medicine, The Scripps Research Institute, Jupiter, Florida, USA. Author Contributions M.D. conceived the project, carried out the experiments, analyzed data, and wrote the paper. L.R., V.V., and L.B. supervised the research and prepared the manuscript. B.S., B.L.M., and A.C. did confocal microscopy and prepared figures. K.K. and G.K. contributed by carrying out cell-based experiments. S.S. designed and contributed to the preparation of Figures 4 and 6. G.G. assisted in the fluorescence polarization experiments. All authors analyzed the results and approved the final version of the manuscript. Funding The work was supported by a grant from the National Research, Development and Innovation Fund of Hungary (K 124045 and FIEK_16-1-2016-0005) and the MedinProt Program of the Hungarian Academy of Sciences (L.B.). L.R.
Funding Information:
and V.V. were supported by a postdoctoral fellowship program by the Hungarian Academy of Sciences. The work of V.V. was supported by a Jańos Bolyai Research Scholarship of the Hungarian Academy of Sciences. Notes The authors declare no competing financial interest.
Publisher Copyright:
Copyright © 2018 American Chemical Society.
Copyright:
Copyright 2019 Elsevier B.V., All rights reserved.
PY - 2018/7/17
Y1 - 2018/7/17
N2 - The nonreceptor tyrosine kinase Src is a central component of the epidermal growth factor (EGF) signaling pathway. Our group recently showed that the Frank-ter Haar syndrome protein Tks4 (tyrosine kinase substrate with four Src homology 3 domains) is also involved in EGF signaling. Here we demonstrate that Tks4 and Src bind directly to each other and elucidate the details of the molecular mechanism of this complex formation. Results of GST pull-down and fluorescence polarization assays show that both a proline-rich SH3 binding motif (PSRPLPDAP, residues 466-474) and an adjacent phosphotyrosine-containing SH2 binding motif (pYEEI, residues 508-511) in Tks4 are responsible for Src binding. These motifs interact with the SH3 and SH2 domains of Src, respectively, leading to a synergistic enhancement of binding strength and a highly stable, "bidentate"-type of interaction. In agreement with these results, we found that the association of Src with Tks4 is permanent and the complex lasts at least 3 h in living cells. We conclude that the interaction of Tks4 with Src may result in the long term stabilization of the kinase in its active conformation, leading to prolonged Src activity following EGF stimulation.
AB - The nonreceptor tyrosine kinase Src is a central component of the epidermal growth factor (EGF) signaling pathway. Our group recently showed that the Frank-ter Haar syndrome protein Tks4 (tyrosine kinase substrate with four Src homology 3 domains) is also involved in EGF signaling. Here we demonstrate that Tks4 and Src bind directly to each other and elucidate the details of the molecular mechanism of this complex formation. Results of GST pull-down and fluorescence polarization assays show that both a proline-rich SH3 binding motif (PSRPLPDAP, residues 466-474) and an adjacent phosphotyrosine-containing SH2 binding motif (pYEEI, residues 508-511) in Tks4 are responsible for Src binding. These motifs interact with the SH3 and SH2 domains of Src, respectively, leading to a synergistic enhancement of binding strength and a highly stable, "bidentate"-type of interaction. In agreement with these results, we found that the association of Src with Tks4 is permanent and the complex lasts at least 3 h in living cells. We conclude that the interaction of Tks4 with Src may result in the long term stabilization of the kinase in its active conformation, leading to prolonged Src activity following EGF stimulation.
UR - http://www.scopus.com/inward/record.url?scp=85049247794&partnerID=8YFLogxK
U2 - 10.1021/acs.biochem.8b00084
DO - 10.1021/acs.biochem.8b00084
M3 - Article
C2 - 29928795
AN - SCOPUS:85049247794
SN - 0006-2960
VL - 57
SP - 4186
EP - 4196
JO - Biochemistry
JF - Biochemistry
IS - 28
ER -