Crystal Structure of the Complex of Human Epidermal Growth Factor and Receptor Extracellular Domains

Ogiso, Hideo; Ishitani, Ryuichiro; Nureki, Osamu; Fukai, Shuya; Yamanaka, Masao; Kim, Jae Hoon; Saito, Kazuki; Sakamoto, Ayako; Inoue, M.; Shirouzu, Mikako; Yokoyama, Shigeyuki

doi:10.1016/s0092-8674(02)00963-7

Cited by 1,023 publications

(1,125 citation statements)

References 59 publications

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“…Dimerization then leads to autophosphorylation of tyrosine residues in the intracellular domain (4)(5)(6). This model has been supported by crystal structures of the extracellular domain of EGFR (7,8) as well as by reports of EGFR dimerization after stimulation with EGF. However, numerous studies have also reported the presence of EGFR dimers or even larger oligomers in the membranes of resting cells (9)(10)(11)(12)(13).…”

Section: Introductionmentioning

confidence: 67%

The Epidermal Growth Factor Receptor Forms Location-Dependent Complexes in Resting Cells

Yavas

Macháň

Wohland

2016

Biophysical Journal

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The epidermal growth factor receptor (EGFR) is a prototypical receptor tyrosine kinase involved in cell growth and proliferation and associated with various cancers. It is commonly assumed that after activation by binding of epidermal growth factor to the extracellular domain it dimerizes, followed by autophosphorylation of tyrosine residues at the intracellular domain. However, its oligomerization state before activation is controversial. In the absence of ligands, EGFR has been found in various, inconsistent amounts of monomeric, inactive dimeric, and oligomeric forms. In addition, evidence suggests that the active conformation is not a simple dimer but contains higher oligomers. As experiments in the past have been conducted at different conditions, we investigate here the influence of cell lines (HEK293, COS-7, and CHO-K1), temperature (room temperature and 37 C), and membrane localization on the quantitation of preformed dimers using SW-FCCS, DC-FCCS, quasi PIE-FCCS, and imaging FCCS. While measurement modality, temperature, and localization on upper or lower membranes have only a limited influence on the dimerization amount observed, the cell line and location to periphery versus center of the cell can change dimerization results significantly. The observed dimerization amount is strongly dependent on the expression level of endogenous EGFR in a cell line and shows a strong cell-to-cell variability even within the same cell line. In addition, using imaging FCCS, we find that dimers have a tendency to be found at the periphery of cells compared to central positions.

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Section: Introductionmentioning

confidence: 67%

The Epidermal Growth Factor Receptor Forms Location-Dependent Complexes in Resting Cells

Yavas

Macháň

Wohland

2016

Biophysical Journal

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“…T-helper cell reactivity against non tolerated rat peptides assists activation of the B cells that will produce antibodies against both rat and hamster HER-2 (Trojan et al, 2001). Moreover, in the nine fragments recognised as the most immunogeneic of the EC and TM domains according to Ogiso et al (2002), six out of 10 substitutions are conservative. In addition, xenogeneic sequences may give rise to 'heteroclitic' peptides that may have a higher affinity for hamster MHC glycoproteins than homologous peptides (Dyall et al, 1998).…”

Section: Discussionmentioning

confidence: 99%

Anti-HER-2 DNA vaccine protects Syrian hamsters against squamous cell carcinomas

et al. 2005

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This paper illustrates the efficacy of DNA vaccination through electroporation in the prevention of oral transplantable carcinoma in Syrian hamsters. At 21 and 7 days before tumour challenge, 19 hamsters were vaccinated with plasmids coding for the extracellular and transmembrane domains of rat HER-2 receptor (EC-TM plasmids), whereas 19 control hamsters were injected intramuscularly with the empty plasmid. Immediately following plasmid injection, hamsters of both groups received two square-wave 25 ms, 375 V cm À1 electric pulses via two electrodes placed on the skin of the injection area. At day 0, all hamsters were challenged in the submucosa of the right cheek pouch with HER-2-positive HCPC I cells established in vitro from an 7,12-dimethylbenz[a]anthraceneinduced oral carcinoma. This challenge gave rise to HER-2-positive buccal neoplastic lesions in 14 controls (73.37%), compared with only seven (36.8%, Po0.0027) vaccinated hamsters. In addition, the vaccinated hamsters displayed both a stronger proliferative and cytotoxic response than the controls and a significant anti-HER-2 antibody response. Most of the hamsters that rejected the challenge displayed the highest antibody titres. These findings suggest that DNA vaccination may have a future in the prevention of HER-2-positive human oral cancer.

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“…EGFR is a transmembrane receptor tyrosine kinase (RTK) and is expressed in multiple cell types (Castillo et al, 2004;Soonthornthumet al, 2011). The molecular structure of EGFR comprises of an extracellular ligand binding domain, a single transmembrane section and an intracellular tyrosine kinase domain (Leahy et al, 2004), which plays an important role in cell proliferation and differentiation upon its activation (Voldborg et al, 1997;Ogiso et al, 2002). EGFR is activated when its cognate ligand epidermal growth factor (EGF) binds to its extracellular domain, which initiates homo or hetero dimerization between the members of the Erb family (Burgess et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

Tyrosine 1045 Codon Mutations in Exon 27 of EGFR are Infrequent in Oral Squamous Cell Carcinomas

Tushar

Ramanathan²

2013

Asian Pacific Journal of Cancer Prevention

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Background: The activation and inactivation of receptor tyrosine kinases are tightly regulated to ensure faithful replication of cells. After having transduced extracellular growth activating signals, activated EGFR is subjected to downregulation either by clathrin mediated endocytosis or c-Cbl mediated proteasome degradation depending on the ligand concentration. c-Cbl is an ubiquitin ligase which requires a phosphorylated tyrosine residue at position 1045 in the cytoplasmic domain of EGFR to interact and add ubiquitin molecules. While activating mutations in exons 19 and 21 have been associated with the development of several cancers, the status of mutations at tyrosine 1045 coding exon 27 of EGFR remain to be investigated. Consistently, defective phosphorylation at 1045 has been associated with sustained phosphorylation of EGFR in non-small lung carcinomas. Hence in the present study we investigated the genetic status of the tyrosine 1045 coding site within exon 27 of EGFR gene to explore for possible occurrence of mutations in this region, especially since no studies have addressed this issue so far. Materials and Methods: Tumor chromosomal DNA isolated from thirty five surgically excised oral squamous cell carcinoma tissues was subjected to PCR amplification with intronic primers flanking the tyrosine 1045 coding exon 27 of EGFR gene. The PCR amplicons were subsequently subjected to direct sequencing to elucidate the mutation status. Results: Sequence analysis identified no mutations in the tyrosine 1045 codon of EGFR in any of the thirty five samples that were analyzed. Conclusions: The lack of identification of mutation in the tyrosine 1045 codon of EGFR suggests that mutations in this region may be relatively rare in oral squamous cell carcinomas. To the best of our knowledge, this study is the first to have explored the genetic status of exon 27 of EGFR in oral squamous cell carcinoma tissue samples.

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Crystal Structure of the Complex of Human Epidermal Growth Factor and Receptor Extracellular Domains

Cited by 1,023 publications

References 59 publications

The Epidermal Growth Factor Receptor Forms Location-Dependent Complexes in Resting Cells

The Epidermal Growth Factor Receptor Forms Location-Dependent Complexes in Resting Cells

Anti-HER-2 DNA vaccine protects Syrian hamsters against squamous cell carcinomas

Tyrosine 1045 Codon Mutations in Exon 27 of EGFR are Infrequent in Oral Squamous Cell Carcinomas

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