2js2 Citations

Specificity determinants of a novel Nck interaction with the juxtamembrane domain of the epidermal growth factor receptor.

Biochemistry 47 3096-108 (2008)
Cited: 11 times
EuropePMC logo PMID: 18269246


Nck is a ubiquitously expressed adaptor protein containing Src homology 2 (SH2) and Src homology 3 (SH3) domains. It integrates downstream effector proteins with cell membrane receptors, such as the epidermal growth factor receptor (EGFR). EGFR plays a critical role in cellular proliferation and differentiation. The 45-residue juxtamembrane domain of EGFR (JM), located between the transmembrane and kinase domains, regulates receptor activation and trafficking to the basolateral membrane of polarized epithelia through a proline-rich motif that resembles a consensus SH3 domain binding site. We demonstrate here that the JM region can bind to Nck, showing a notable binding preference for the second SH3 domain. To elucidate the structural determinants for this interaction, we have determined the NMR solution structures of both the first and second Nck SH3 domains (Nck1-1 and Nck1-2). These domains adopt a canonical SH3 beta-barrel-like fold, containing five antiparallel strands separated by three loop regions and one 3 10-helical turn. Chemical shift perturbation studies have identified the residues that form the binding cleft of Nck1-2, which are primarily located in the RT and n-Src loops. JM binds to Nck1-2 with an affinity of approximately 80 microM through a positively charged sequence near the N-terminus, as opposed to the polyproline sequence. The two Nck SH3 domains exhibit both steric and electrostatic differences in their RT-Src and n-Src loops, and a model of the Nck1-2 domain complexed with the JM highlights the factors that define the putative binding mode for this ligand.

Reviews citing this publication (2)

  1. Calmodulin-mediated regulation of the epidermal growth factor receptor. Sánchez-González P, Jellali K, Villalobo A. FEBS J. 277 327-342 (2010)
  2. Calmodulin-mediated regulation of the epidermal growth factor receptor. Sánchez-González P, Jellali K, Villalobo A. FEBS J. 277 327-342 (2010)

Articles citing this publication (9)

  1. Nck adapter proteins: functional versatility in T cells. Lettau M, Pieper J, Janssen O. Cell Commun. Signal 7 1 (2009)
  2. Proline isomerization preorganizes the Itk SH2 domain for binding to the Itk SH3 domain. Severin A, Joseph RE, Boyken S, Fulton DB, Andreotti AH. J. Mol. Biol. 387 726-743 (2009)
  3. Direct regulation of nephrin tyrosine phosphorylation by Nck adaptor proteins. New LA, Keyvani Chahi A, Jones N. J. Biol. Chem. 288 1500-1510 (2013)
  4. Transmembrane helix orientation influences membrane binding of the intracellular juxtamembrane domain in Neu receptor peptides. Matsushita C, Tamagaki H, Miyazawa Y, Aimoto S, Smith SO, Sato T. Proc. Natl. Acad. Sci. U.S.A. 110 1646-1651 (2013)
  5. Autosomal recessive polycystic kidney disease epithelial cell model reveals multiple basolateral epidermal growth factor receptor sorting pathways. Ryan S, Verghese S, Cianciola NL, Cotton CU, Carlin CR. Mol. Biol. Cell 21 2732-2745 (2010)
  6. Automated assignment of NMR chemical shifts using peak-particle dynamics simulation with the DYNASSIGN algorithm. Schmucki R, Yokoyama S, Güntert P. J. Biomol. NMR 43 97-109 (2009)
  7. c-ABL modulates MAP kinases activation downstream of VEGFR-2 signaling by direct phosphorylation of the adaptor proteins GRB2 and NCK1. Anselmi F, Orlandini M, Rocchigiani M, De Clemente C, Salameh A, Lentucci C, Oliviero S, Galvagni F. Angiogenesis 15 187-197 (2012)
  8. Autoinhibitory interaction in the multidomain adaptor protein Nck: possible roles in improving specificity and functional diversity. Takeuchi K, Sun ZY, Park S, Wagner G. Biochemistry 49 5634-5641 (2010)
  9. Coordinated activation of the Rac-GAP β2-chimaerin by an atypical proline-rich domain and diacylglycerol. Gutierrez-Uzquiza A, Colon-Gonzalez F, Leonard TA, Canagarajah BJ, Wang H, Mayer BJ, Hurley JH, Kazanietz MG. Nat Commun 4 1849 (2013)