In vivo identification of the interaction site of ErbB2 extracellular domain with its autoinhibitor

Hu, Pinliang; Feng, Jie; Zhou, Tao; Wang, Jianbo; Bao-feng, Jing; Yu, Ming; Hu, Meiru; Zhang, Xuemin; Shen, Beifen; Guo, Ning

doi:10.1002/jcp.20409

Cited by 16 publications

(10 citation statements)

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“…2001; Wong 2003). Functional compensation between ErbB2 and ErbB1 has been directly demonstrated in vitro by the induction of ErbB1 expression and activation of ErbB1 signaling in response to loss of ErbB2 function (Hu et al . 2005b).…”

Section: Discussionmentioning

confidence: 99%

Requirement for ErbB2/ErbB signaling in developing cartilage and bone

et al. 2007

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During endochondral ossification, the skeletal elements of vertebrate limbs form and elongate via coordinated control of chondrocyte and osteoblast differentiation and proliferation. The role of signaling by the ErbB family of receptor tyrosine kinases, which consists of ErbB1 (epidermal growth factor receptor or EGFR), ErbB2, ErbB3 and ErbB4, has been little studied during cartilage and bone development. Signaling by the ErbB network generates a diverse array of cellular responses via formation of ErbB dimers activated by distinct ligands that produce distinct signal outputs. Herstatin is a soluble ErbB2 receptor that acts in a dominant negative fashion to inhibit ErbB signaling by binding to endogenous ErbB receptors, preventing functional dimer formation. Here, we examine the effects of Herstatin on limb skeletal element development in transgenic mice, achieved via Prx1 promoter-driven expression in limb cartilage and bone. The limb skeletal elements of Prx1-Herstatin embryos are shortened, and chondrocyte maturation and osteoblast differentiation are delayed. In addition, proliferation by chondrocytes and periosteal cells of Prx1-Herstatin limb skeletal elements is markedly reduced. Our study identifies requirements for ErbB signaling in the maintenance of chondrocyte and osteoblast proliferation involved in the timely progression of chondrocyte maturation and periosteal osteoblast differentiation.

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Section: Discussionmentioning

confidence: 99%

Requirement for ErbB2/ErbB signaling in developing cartilage and bone

et al. 2007

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“…The binding site of herstatin of the ErbB2 ECD domain was proposed to be at the S1 domain (here domain II). That observation was verified by inmunoprecipitation, confocal microscopy and fluorescence resonance energy transfer experiments [29]. Very recently, Fuentes et al have published a 20 ns MD study and a fluctuation analysis of the interaction between ErbB2 and a combination of trastuzumab and pertuzumab antibodies [30].…”

Section: Introductionmentioning

confidence: 87%

Conformational flexibility of the ErbB2 ectodomain and trastuzumab antibody complex as revealed by molecular dynamics and principal component analysis

et al. 2012

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Human epidermal growth factor receptor 2 (ErbB2) is a transmembrane oncoprotein that is over expressed in breast cancer. A successful therapeutic treatment is a monoclonal antibody called trastuzumab which interacts with the ErbB2 extracellular domain (ErbB2-ECD). A better understanding of the detailed structure of the receptor-antibody interaction is indeed of prime interest for the design of more effective anticancer therapies. In order to discuss the flexibility of the complex ErbB2-ECD/ trastuzumab, we present, in this study, a multi-nanosecond molecular dynamics simulation (MD) together with an analysis of fluctuations, through a principal component analysis (PCA) of this system. Previous to this step and in order to validate the simulations, we have performed a detailed analysis of the variable antibody domain interactions with the extracellular domain IV of ErbB2. This structure has been statically elucidated by x-ray studies. Indeed, the simulation results are in excellent agreement with the available experimental information during the full trajectory. The PCA shows eigenvector fluctuations resulting in a hinge motion in which domain II and C H domains approach each other. This move is likely stabilized by the formation of H-bonds and salt bridge interactions between residues of the dimerization arm in the domain II and trastuzumab residues located in the C H domain. Finally, we discuss the flexibility of the MD/PCA model in relation with the static x-ray structure. A movement of the antibody toward the dimerization domain of the ErbB2 receptor is reported for the first time. This finding could have important consequences on the biological action of the monoclonal antibody.Keywords Extracellular ErbB2 receptor . Herceptin . Molecular dynamics . Principal component analysis . Trastuzumab IntroductionThe human epidermal growth factor receptors (EGFR) HER1 (ErbB1, EGFR), HER2 (ErbB2), HER3 (ErbB3) and HER4 (ErbB4) belong to the family of receptor tyrosine kinase proteins. These receptors are engaged in the regulation of many processes such as cell proliferation, differentiation and apoptosis. Loss of regulation of these receptors has a great impact in a number of human diseases, such as cancer [1,2]. All EGFR receptors contain three different regions: an extracellular (ectodomain, ECD) ligand-binding region, a single membrane-spanning domain and a cytoplasmatic tyrosine kinase domain. The extracellular EGFR domains have been crystallographically elucidated by several research groups [3][4][5][6][7][8][9][10]. The x-ray structure-based models show the appearance of two large homologous domains (L) and two cysteinerich domains (CR), in the order L1-CR1-L2-CR2 which is simply known as I-II-III-IV domains, (see Scheme 1).Electronic supplementary material The online version of this article

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“…The dimeric structure of the growth factor receptor ErbB2 consists of three conservative segments: the extracellular domain, the transmembrane region, and the tyrosine-kinase region (Hu et al, 2005). The molecular chaperone Hsp90 can lead to the phosphorylation of the intracellular tyrosine-kinase by binding to the extracellular domain of ErbB2 receptor, and then activating downstream PI3K/Akt and ERK1/2 pathways.…”

Section: Discussionmentioning

confidence: 99%

Heat shock protein 90 protects rat mesenchymal stem cells against hypoxia and serum deprivation-induced apoptosis via the PI3K/Akt and ERK1/2 pathways

Gao

Xie

et al. 2010

J. Zhejiang Univ. Sci. B

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Mesenchymal stem cell (MSC) transplantation has shown a therapeutic potential to repair the ischemic and infracted myocardium, but the effects are limited by the apoptosis and loss of donor cells in host cardiac microenvironment. The aim of this study is to explore the cytoprotection of heat shock protein 90 (Hsp90) against hypoxia and serum deprivation-induced apoptosis and the possible mechanisms in rat MSCs. Cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Apoptosis was assessed by Hoechst 33258 nuclear staining and flow cytometric analysis with annexin V/PI staining. The gene expression of Toll-like receptor-4 (TLR-4) and V-erb-b2 erythroblastic leukemia viral oncogene homolog 2 (ErbB2) was detected by real-time polymerase chain reaction (PCR). The protein levels of cleaved caspase-3, Bcl-2, Bcl-xL, Bax, total-ERK, phospho-ERK, total-Akt, phospho-Akt, and Hsp90 were detected by Western blot. The production of nitric oxide was measured by spectrophotometric assay. Hsp90 improves MSC viability and protects MSCs against apoptosis induced by serum deprivation and hypoxia. The protective role of Hsp90 not only elevates Bcl-2/Bax and Bcl-xL/Bax expression and attenuates cleaved caspase-3 expression via down-regulating membrane TLR-4 and ErbB2 receptors and then activating their downstream PI3K/Akt and ERK1/2 pathways, but also enhances the paracrine effect of MSCs. These findings demonstrated a novel and effective treatment strategy against MSC apoptosis in cell transplantation.

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In vivo identification of the interaction site of ErbB2 extracellular domain with its autoinhibitor

Cited by 16 publications

References 55 publications

Requirement for ErbB2/ErbB signaling in developing cartilage and bone

Requirement for ErbB2/ErbB signaling in developing cartilage and bone

Conformational flexibility of the ErbB2 ectodomain and trastuzumab antibody complex as revealed by molecular dynamics and principal component analysis

Heat shock protein 90 protects rat mesenchymal stem cells against hypoxia and serum deprivation-induced apoptosis via the PI3K/Akt and ERK1/2 pathways

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