Molecular architecture of the ErbB2 extracellular domain homodimer

Hu, Shi; Sun, Yuna; Meng, Yanchun; Wang, Xiaoze; Yang, Weili; Fu, Wenyan; Guo, Haipeng; Qian, Weizhu; Hou, Sheng; Li, Bohua; Rao, Zihe; Lou, Zhiyong; Guo, Yajun

doi:10.18632/oncotarget.2713

Cited by 47 publications

(40 citation statements)

References 32 publications

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“…Recently, we have determined the crystal structure of ErbB2 in complex with H2-18. Our crystallographic analysis confirmed that the epitope of H2-18 is in domain I of ErbB2 [23]. Competitive binding assays also showed that H2-18 did not compete with either trastuzumab or pertuzumab for binding to ErbB2 (Supplementary Figure S1).…”

Section: Resultssupporting

confidence: 71%

An anti-ErbB2 fully human antibody circumvents trastuzumab resistance

Wang

Zhang

et al. 2016

Oncotarget

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Trastuzumab, an anti-HER2/ErbB2 humanized antibody, has shown great clinical benefits in ErbB2-positive breast cancer treatment. Despite of its effectiveness, response rate to trastuzumab is limited and resistance is common. Here, we developed a new anti-ErbB2 antibody, denoted as H2-18, which was isolated from a phage display human antibody library. Previous studies have demonstrated that trastuzumab recognizes the juxtamembrane region of domain IV, and pertuzumab, another humanized ErbB2-specific antibody, binds to ErbB2 near the center of domain II. Our crystallographic analysis showed that the epitope recognized by H2-18 is within domain I of the ErbB2 molecule. H2-18 potently induced programmed cell death (PCD) in both trastuzumab-sensitive and -resistant breast cancer cell lines, while trastuzumab and pertuzumab, either used alone or in combination, only exhibits very weak PCD-inducing activity. More importantly, H2-18 could inhibit the growth of trastuzumab-resistant breast cancer cells far more effectively than trastuzumab plus pertuzumab, both in vitro and in vivo. In conclusion, H2-18 shows a unique ability to overcome trastuzumab resistance, suggesting that it has the great potential to be translated to the clinic.

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Section: Resultssupporting

confidence: 71%

An anti-ErbB2 fully human antibody circumvents trastuzumab resistance

Wang

Zhang

et al. 2016

Oncotarget

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“…Another potential explanation for negative cooperativity could be steric hindrance for the second HER2 molecule. Due to the high flexibility of the domain IV of HER2, a total of 55 residues on the C-terminus could not be resolved, which is also the case for other structures of HER2 (Franklin et al, 2004;Hu et al, 2015). However, the HER2 molecule of the trastuzumab Fab-HER2 complex structure (Cho et al, 2003), where the Fab fragment binds at the lower end of domain IV, shows more electron density in domain IV and therefore misses only 24 residues on the C-terminal part.…”

Section: Discussionmentioning

confidence: 99%

Fcab-HER2 Interaction: a Ménage à Trois. Lessons from X-Ray and Solution Studies

et al. 2017

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The crystallizable fragment (Fc) of the immunoglobulin class G (IgG) is an attractive scaffold for the design of novel therapeutics. Upon engineering the C-terminal loops in the CH3 domains, Fcabs (Fc domain with antigen-binding sites) can be designed. We present the first crystal structures of Fcabs, i.e., of the HER2-binding clone H10-03-6 having the AB and EF loop engineered and the stabilized version STAB19 derived by directed evolution. Comparison with the crystal structure of the Fc wild-type protein reveals conservation of the overall domain structures but significant differences in EF-loop conformations. Furthermore, we present the first Fcab-antigen complex structures demonstrating the interaction between the engineered Fcab loops with domain IV of HER2. The crystal structures of the STAB19-HER2 and H10-03-6-HER2 complexes together with analyses by isothermal titration calorimetry, SEC-MALS, and fluorescence correlation spectroscopy show that one homodimeric Fcab binds two HER2 molecules following a negative cooperative binding behavior.

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“…Reflecting our previous in vitro computational model, we account for binding of matrix-bound ligands to VEGFR2 (previously demonstrated [18, 19]) and VEGFR1 (assumed to occur). We assume that endothelial basement membrane-bound growth factor within 25nm of the cell surface is accessible to cell surface receptors, based on the length of the extracellular domain of the related RTKs ErbB2 and ErbB3 (11.3–16.4nm) [73–75], and assuming some flexibility in cell position and shape. We calculated the resulting fraction of EBM accessible to cell surface receptors (S7 Table), and scaled the corresponding reaction on-rates (Table 2, see S1 Equations).…”

Section: Methodsmentioning

confidence: 99%

A computational analysis of in vivo VEGFR activation by multiple co-expressed ligands

Clegg

Gabhann

2017

PLoS Comput Biol

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The splice isoforms of vascular endothelial growth A (VEGF) each have different affinities for the extracellular matrix (ECM) and the coreceptor NRP1, which leads to distinct vascular phenotypes in model systems expressing only a single VEGF isoform. ECM-immobilized VEGF can bind to and activate VEGF receptor 2 (VEGFR2) directly, with a different pattern of site-specific phosphorylation than diffusible VEGF. To date, the way in which ECM binding alters the distribution of isoforms of VEGF and of the related placental growth factor (PlGF) in the body and resulting angiogenic signaling is not well-understood. Here, we extend our previous validated cell-level computational model of VEGFR2 ligation, intracellular trafficking, and site-specific phosphorylation, which captured differences in signaling by soluble and immobilized VEGF, to a multi-scale whole-body framework. This computational systems pharmacology model captures the ability of the ECM to regulate isoform-specific growth factor distribution distinctly for VEGF and PlGF, and to buffer free VEGF and PlGF levels in tissue. We show that binding of immobilized growth factor to VEGF receptors, both on endothelial cells and soluble VEGFR1, is likely important to signaling in vivo. Additionally, our model predicts that VEGF isoform-specific properties lead to distinct profiles of VEGFR1 and VEGFR2 binding and VEGFR2 site-specific phosphorylation in vivo, mediated by Neuropilin-1. These predicted signaling changes mirror those observed in murine systems expressing single VEGF isoforms. Simulations predict that, contrary to the ‘ligand-shifting hypothesis,’ VEGF and PlGF do not compete for receptor binding at physiological concentrations, though PlGF is predicted to slightly increase VEGFR2 phosphorylation when over-expressed by 10-fold. These results are critical to design of appropriate therapeutic strategies to control VEGF availability and signaling in regenerative medicine applications.

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Molecular architecture of the ErbB2 extracellular domain homodimer

Cited by 47 publications

References 32 publications

An anti-ErbB2 fully human antibody circumvents trastuzumab resistance

An anti-ErbB2 fully human antibody circumvents trastuzumab resistance

Fcab-HER2 Interaction: a Ménage à Trois. Lessons from X-Ray and Solution Studies

A computational analysis of in vivo VEGFR activation by multiple co-expressed ligands

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