Spatial Structure of the Transmembrane Domain Heterodimer of ErbB1 and ErbB2 Receptor Tyrosine Kinases

Mineev, Konstantin S.; Bocharov, Eduard V.; Pustovalova, Yulia; Bocharova, Olga V.; Chupin, Vladimir; Arseniev, Alexander S.

doi:10.1016/j.jmb.2010.05.016

Cited by 120 publications

(119 citation statements)

References 64 publications

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“…The EGFR transmembrane domain has been previously described to be critically involved in the communication between the ECD and the intracellular TKD. Therefore, the possible regulatory function of the TMD has received substantial attention (2,8,(33)(34)(35)(36)(37)(38). Two GxxxG dimerization motifs present on both ends of the TMD are supposed to regulate the association of ligand-free and ligandbound dimers (2,8).…”

Section: Resultsmentioning

confidence: 99%

N -Glycosylation as determinant of epidermal growth factor receptor conformation in membranes

Kaszuba

Grzybek

Orłowski

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

146

156

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The epidermal growth factor receptor (EGFR) regulates several critical cellular processes and is an important target for cancer therapy. In lieu of a crystallographic structure of the complete receptor, atomistic molecular dynamics (MD) simulations have recently shown that they can excel in studies of the full-length receptor. Here we present atomistic MD simulations of the monomeric N-glycosylated human EGFR in biomimetic lipid bilayers that are, in parallel, also used for the reconstitution of full-length receptors. This combination enabled us to experimentally validate our simulations, using ligand binding assays and antibodies to monitor the conformational properties of the receptor reconstituted into membranes. We find that N-glycosylation is a critical determinant of EGFR conformation, and specifically the orientation of the EGFR ectodomain relative to the membrane. In the absence of a structure for full-length, posttranslationally modified membrane receptors, our approach offers new means to structurally define and experimentally validate functional properties of cell surface receptors in biomimetic membrane environments.R eceptor tyrosine kinases (RTKs) are cell surface receptors that receive and transduce signals mediating a variety of critical cellular processes, including cell growth, migration, proliferation, differentiation, and apoptosis. Among the many RTKs, the most studied is the epidermal growth factor receptor (EGFR), not least because of its involvement in the development and progression of epidermoid cancers and its resulting importance as a target for antineoplastic therapies.Structurally, the EGFR consists of the ectodomain (ECD) (further subdivided into four subdomains, DI-IV), the transmembrane domain (TMD), and the intracellular tyrosine kinase domain (TKD). Ligand binding induces conformational transitions of the ECD that stabilize receptor dimerization, culminating in the activation of the intracellular TKD and subsequent propagation of the activation signal (1). To prevent receptor activation and signaling in the absence of ligand, the structurally tethered ECD of monomeric EGFR blocks the intrinsic capacity of the TMD and the intracellular TKD to dimerize (2). Ligand binding is believed to release the self-inhibitory tether and facilitate receptor oligomerization and activation (3-6). A detailed understanding of the structural regulation of the intact full-length receptors in their native membranes promises to reveal the molecular basis for receptor regulation (7); however, the methodological limitations associated with crystallizing transmembrane proteins, together with the high flexibility of the full-length receptor, have prevented high-resolution crystallographic analysis.To fill this gap, extensive atom-scale molecular dynamics (MD) simulations were recently performed to elucidate the structural dynamics of the EGFR in a two-component lipid bilayer (8). These studies suggest a large interfacial contact area between the membrane and the ecto-and intracellular domains of the unli...

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Section: Resultsmentioning

confidence: 99%

N -Glycosylation as determinant of epidermal growth factor receptor conformation in membranes

Kaszuba

Grzybek

Orłowski

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

146

156

View full text Add to dashboard Cite

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“…ErbB receptors have two TMD-dimerization motifs, and bisulfide crosslinking and NMR studies showed that ErbB TMDs predominantly interact through their N-terminal GG4-like motif. Interestingly, because this dimerization occurs in an angle of 4665˚, such dimers might create a wedge-like structure in the plasma membrane (Lu et al, 2010;Mineev et al, 2010). Clustering of such wedge-like structures could potentially induce a concave membrane curvature.…”

mentioning

confidence: 99%

EGFR endocytosis requires its kinase activity and N-terminal transmembrane dimerization motif

Heukers

Vermeulen

Fereidouni

et al. 2013

Journal of Cell Science

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Summary EGFR signaling is attenuated by endocytosis and degradation of receptor-ligand complexes in lysosomes. Endocytosis of EGFR is known to be regulated by multiple post-translational modifications. The observation that prevention of these modifications does not block endocytosis completely, suggests the involvement of other mechanism(s). Recently, receptor clustering has been suggested to induce internalization of multiple types of membrane receptors. However, the mechanism of clustering-induced internalization remains unknown. We have used biparatopic antibody fragments from llama (VHHs) to induce EGFR clustering without stimulating tyrosine kinase activity. Using this approach, we have found an essential role for the N-terminal GG4-like dimerization motif in the transmembrane domain (TMD) for clustering-induced internalization. Moreover, conventional EGF-induced receptor internalization depends exclusively on this TMD dimerization and kinase activity. Mutations in this dimerization motif eventually lead to reduced EGFR degradation and sustained signaling. We propose a novel role for the TMD dimerization motif in the negative-feedback control of EGFR. The widely conserved nature of GG4-like dimerization motifs in transmembrane proteins suggests a general role for these motifs in clustering-induced internalization.

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“…Recent structural studies on the isolated TM helices of ErbB2, EphA1, EphA2, ErbB3, and the heterodimer ErbB1/ ErbB2 showed that the TM helices, at least in these systems, dimerize in two different ways: either right-handed parallel or left-handed coiled-coil ␣-helical dimers (12)(13)(14)(15)(16)(17). Ambiguity remains about what governs the arrangement in either of the two conformations or whether both conformations are relevant for activation and may in fact represent different activation states for a given RTK dimer.…”

mentioning

confidence: 99%

Hydrophobic Matching Controls the Tilt and Stability of the Dimeric Platelet-derived Growth Factor Receptor (PDGFR) β Transmembrane Segment

Muhle‐Goll

Hoffmann

Afonin

et al. 2012

Journal of Biological Chemistry

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Background: Dimerization regulates activation of PDGF receptor in signal transduction. Results: The transmembrane segment of PDGFR forms a left-handed helical dimer, which becomes more tilted and less stable in model membranes with decreasing lipid acyl chain lengths. Conclusion:The membrane thickness controls the ability of the transmembrane segments to dimerize. Significance: Receptor dimerization and activation in vivo may require relocation to thick lipid rafts.

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Spatial Structure of the Transmembrane Domain Heterodimer of ErbB1 and ErbB2 Receptor Tyrosine Kinases

Cited by 120 publications

References 64 publications

N -Glycosylation as determinant of epidermal growth factor receptor conformation in membranes

N -Glycosylation as determinant of epidermal growth factor receptor conformation in membranes

EGFR endocytosis requires its kinase activity and N-terminal transmembrane dimerization motif

Hydrophobic Matching Controls the Tilt and Stability of the Dimeric Platelet-derived Growth Factor Receptor (PDGFR) β Transmembrane Segment

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