Bipartite Tetracysteine Display Reveals Allosteric Control of Ligand-Specific EGFR Activation

Scheck, Rebecca A.; Lowder, Melissa A.; Appelbaum, Jacob; Schepartz, Alanna

doi:10.1021/cb300216f

Cited by 57 publications

(123 citation statements)

References 42 publications

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“…Intriguingly, this region appears to form an antiparallel helical dimer in the NMR-guided structural model of the TM domain plus iJM helix in Figure 3C (Endres et al 2013). A chemical biology approach using bipartite tetracysteine display has also provided evidence for formation of this antiparallel helical dimer in the intact receptor in cells (Scheck et al 2012). Formation of such a dimer offers a way to break the artifactual "daisy chain" seen in crystals of the isolated EGFR TKD, in which the iJM region of each molecule cradles the carboxy lobe of its neighbor, and each TKD simultaneously acts as activator and receiver.…”

Section: The Intracellular Juxtamembrane (Ijm) Regionmentioning

confidence: 90%

The EGFR Family: Not So Prototypical Receptor Tyrosine Kinases

Lemmon

Schlessinger

Ferguson

2014

Cold Spring Harbor Perspectives in Biology

367

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The epidermal growth factor receptor (EGFR) was among the first receptor tyrosine kinases (RTKs) for which ligand binding was studied and for which the importance of ligand-induced dimerization was established. As a result, EGFR and its relatives have frequently been termed "prototypical" RTKs. Many years of mechanistic studies, however, have revealed that-far from being prototypical-the EGFR family is quite unique. As we discuss in this review, the EGFR family uses a distinctive "receptor-mediated" dimerization mechanism, with ligand binding inducing a dramatic conformational change that exposes a dimerization arm. Intracellular kinase domain regulation in this family is also unique, being driven by allosteric changes induced by asymmetric dimer formation rather than the more typical activationloop phosphorylation. EGFR family members also distinguish themselves from other RTKs in having an intracellular juxtamembrane (JM) domain that activates (rather than autoinhibits) the receptor and a very large carboxy-terminal tail that contains autophosphorylation sites and serves an autoregulatory function. We discuss recent advances in mechanistic aspects of all of these components of EGFR family members, attempting to integrate them into a view of how RTKs in this important class are regulated at the cell surface.

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Section: The Intracellular Juxtamembrane (Ijm) Regionmentioning

confidence: 90%

The EGFR Family: Not So Prototypical Receptor Tyrosine Kinases

Lemmon

Schlessinger

Ferguson

2014

Cold Spring Harbor Perspectives in Biology

367

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“…The juxtamembrane segment (JM) is a short (37 aa) sequence that links the extracellular ligand binding and transmembrane domains to the intracellular kinase domain and stabilizes the receptor active state (Jura et al, 2009a). We discovered that binding of the growth factor EGF to the EGFR extracellular domain induced the formation of a discrete anti-parallel coiled coil (Jura et al, 2009a) within the juxtamembrane-A (JM-A) segment, whereas binding of the alternative growth factor TGF-α induced an alternative, helical interface whose structure was not established (Figure 1A) (Scheck et al, 2012). As predicted by NOE’s seen in short peptide models (Jura et al, 2009a), the EGF-induced antiparallel structure is characterized by leucine residues at the a and d positions of the paired heptad repeat and complementary electrostatic interactions at positions e and g (Figure S1B).…”

Section: Introductionmentioning

confidence: 99%

Growth Factor Identity Is Encoded by Discrete Coiled-Coil Rotamers in the EGFR Juxtamembrane Region

Doerner

Scheck

Schepartz

2015

Chemistry & Biology

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Summary Binding of the growth factor TGF-α to the EGFR extracellular domain is encoded through the formation of a unique anti-parallel coiled coil within the juxtamembrane segment. This new coiled coil is an ‘inside-out’ version of the coiled coil formed in the presence of EGF. A third, intermediary coiled coil interface is formed in the juxtamembrane segment when EGFR is stimulated with betacellulin. The seven growth factors that activate EGFR in mammalian systems (EGF, TGF-α, epigen, epiregulin, betacellulin, heparin-binding EGF, and amphiregulin) fall into distinct categories in which the structure of the coiled coil induced within the juxtamembrane segment correlates with cell state. The observation that coiled coil state tracks with the downstream signaling profiles for each ligand provides evidence for growth factor functional selectivity by EGFR. Encoding growth factor identity in alternative coiled coil rotamers provides a simple and elegant method for communicating chemical information across the plasma membrane.

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“…1A). A key step in switching on the receptor is the formation of an asymmetric dimer of the kinase domains, in which one kinase, termed the activator, allosterically activates the other one, termed the receiver (12)(13)(14)(15)(16)(17). The transmembrane helices of the receptors also dimerize (18)(19)(20).…”

mentioning

confidence: 99%

Analysis of the Role of the C-Terminal Tail in the Regulation of the Epidermal Growth Factor Receptor

Kovács

Das

Wang

et al. 2015

Molecular and Cellular Biology

100

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The ϳ230-residue C-terminal tail of the epidermal growth factor receptor (EGFR) is phosphorylated upon activation. We examined whether this phosphorylation is affected by deletions within the tail and whether the two tails in the asymmetric active EGFR dimer are phosphorylated differently. We monitored autophosphorylation in cells using flow cytometry and found that the first ϳ80 residues of the tail are inhibitory, as demonstrated previously. The entire ϳ80-residue span is important for autoinhibition and needs to be released from both kinases that form the dimer. These results are interpreted in terms of crystal structures of the inactive kinase domain, including two new ones presented here. Deletions in the remaining portion of the tail do not affect autophosphorylation, except for a six-residue segment spanning Tyr 1086 that is critical for activation loop phosphorylation. Phosphorylation of the two tails in the dimer is asymmetric, with the activator tail being phosphorylated somewhat more strongly. Unexpectedly, we found that reconstitution of the transmembrane and cytoplasmic domains of EGFR in vesicles leads to a peculiar phenomenon in which kinase domains appear to be trapped between stacks of lipid bilayers. This artifactual trapping of kinases between membranes enhances an intrinsic functional asymmetry in the two tails in a dimer.

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Bipartite Tetracysteine Display Reveals Allosteric Control of Ligand-Specific EGFR Activation

Cited by 57 publications

References 42 publications

The EGFR Family: Not So Prototypical Receptor Tyrosine Kinases

The EGFR Family: Not So Prototypical Receptor Tyrosine Kinases

Growth Factor Identity Is Encoded by Discrete Coiled-Coil Rotamers in the EGFR Juxtamembrane Region

Analysis of the Role of the C-Terminal Tail in the Regulation of the Epidermal Growth Factor Receptor

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