Glioma Specific Extracellular Missense Mutations in the First Cysteine Rich Region of Epidermal Growth Factor Receptor (EGFR) Initiate Ligand Independent Activation
The epidermal growth factor receptor (EGFR) is overexpressed or mutated in glioma. Recently, a series of missense mutations in the extracellular domain (ECD) of EGFR were reported in glioma patients. Some of these mutations clustered within a cysteine-rich region of the EGFR targeted by the therapeu...
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| Format: | Article |
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MDPI AG
2011-04-01
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| Series: | Cancers |
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| Online Access: | http://www.mdpi.com/2072-6694/3/2/2032/ |
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| author | V. Chandana Epa Andrew M. Scott Timothy E. Adams Diana X. Cao Sameer A. Greenall Jacqui F. Donoghue Anna Cvrljevic Terrance G. Johns Susie I. Ymer |
| author_facet | V. Chandana Epa Andrew M. Scott Timothy E. Adams Diana X. Cao Sameer A. Greenall Jacqui F. Donoghue Anna Cvrljevic Terrance G. Johns Susie I. Ymer |
| author_sort | V. Chandana Epa |
| collection | DOAJ |
| description | The epidermal growth factor receptor (EGFR) is overexpressed or mutated in glioma. Recently, a series of missense mutations in the extracellular domain (ECD) of EGFR were reported in glioma patients. Some of these mutations clustered within a cysteine-rich region of the EGFR targeted by the therapeutic antibody mAb806. This region is only exposed when EGFR activates and appears to locally misfold during activation. We expressed two of these mutations (R324L and E330K) in NR6 mouse fibroblasts, as they do not express any EGFR-related receptors. Both mutants were autophosphorylated in the absence of ligand and enhanced cell survival and anchorage-independent and xenograft growth. The ECD truncation that produces the de2-7EGFR (or EGFRvIII), the most common EGFR mutation in glioma, generates a free cysteine in this same region. Using a technique optimized for detecting disulfide-bonded dimers, we definitively demonstrated that the de2-7EGFR is robustly dimerized and that ablation of the free cysteine prevents dimerization and activation. Modeling of the R324L mutation suggests it may cause transient breaking of disulfide bonds, leading to similar disulfide-bonded dimers as seen for the de2-7EGFR. These ECD mutations confirm that the cysteine-rich region of EGFR around the mAb806 epitope has a significant role in receptor activation. |
| first_indexed | 2024-03-12T06:02:23Z |
| format | Article |
| id | doaj.art-f5291be214a648d49dd08eea4d5800ed |
| institution | Directory Open Access Journal |
| issn | 2072-6694 |
| language | English |
| last_indexed | 2024-03-12T06:02:23Z |
| publishDate | 2011-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cancers |
| spelling | doaj.art-f5291be214a648d49dd08eea4d5800ed2023-09-03T04:04:46ZengMDPI AGCancers2072-66942011-04-01322032204910.3390/cancers3022032Glioma Specific Extracellular Missense Mutations in the First Cysteine Rich Region of Epidermal Growth Factor Receptor (EGFR) Initiate Ligand Independent ActivationV. Chandana EpaAndrew M. ScottTimothy E. AdamsDiana X. CaoSameer A. GreenallJacqui F. DonoghueAnna CvrljevicTerrance G. JohnsSusie I. YmerThe epidermal growth factor receptor (EGFR) is overexpressed or mutated in glioma. Recently, a series of missense mutations in the extracellular domain (ECD) of EGFR were reported in glioma patients. Some of these mutations clustered within a cysteine-rich region of the EGFR targeted by the therapeutic antibody mAb806. This region is only exposed when EGFR activates and appears to locally misfold during activation. We expressed two of these mutations (R324L and E330K) in NR6 mouse fibroblasts, as they do not express any EGFR-related receptors. Both mutants were autophosphorylated in the absence of ligand and enhanced cell survival and anchorage-independent and xenograft growth. The ECD truncation that produces the de2-7EGFR (or EGFRvIII), the most common EGFR mutation in glioma, generates a free cysteine in this same region. Using a technique optimized for detecting disulfide-bonded dimers, we definitively demonstrated that the de2-7EGFR is robustly dimerized and that ablation of the free cysteine prevents dimerization and activation. Modeling of the R324L mutation suggests it may cause transient breaking of disulfide bonds, leading to similar disulfide-bonded dimers as seen for the de2-7EGFR. These ECD mutations confirm that the cysteine-rich region of EGFR around the mAb806 epitope has a significant role in receptor activation.http://www.mdpi.com/2072-6694/3/2/2032/EGFRde2-7EGFRextracellular domain mutationautoactivationdimerizationdisulfide bondfree cysteine |
| spellingShingle | V. Chandana Epa Andrew M. Scott Timothy E. Adams Diana X. Cao Sameer A. Greenall Jacqui F. Donoghue Anna Cvrljevic Terrance G. Johns Susie I. Ymer Glioma Specific Extracellular Missense Mutations in the First Cysteine Rich Region of Epidermal Growth Factor Receptor (EGFR) Initiate Ligand Independent Activation Cancers EGFR de2-7EGFR extracellular domain mutation autoactivation dimerization disulfide bond free cysteine |
| title | Glioma Specific Extracellular Missense Mutations in the First Cysteine Rich Region of Epidermal Growth Factor Receptor (EGFR) Initiate Ligand Independent Activation |
| title_full | Glioma Specific Extracellular Missense Mutations in the First Cysteine Rich Region of Epidermal Growth Factor Receptor (EGFR) Initiate Ligand Independent Activation |
| title_fullStr | Glioma Specific Extracellular Missense Mutations in the First Cysteine Rich Region of Epidermal Growth Factor Receptor (EGFR) Initiate Ligand Independent Activation |
| title_full_unstemmed | Glioma Specific Extracellular Missense Mutations in the First Cysteine Rich Region of Epidermal Growth Factor Receptor (EGFR) Initiate Ligand Independent Activation |
| title_short | Glioma Specific Extracellular Missense Mutations in the First Cysteine Rich Region of Epidermal Growth Factor Receptor (EGFR) Initiate Ligand Independent Activation |
| title_sort | glioma specific extracellular missense mutations in the first cysteine rich region of epidermal growth factor receptor egfr initiate ligand independent activation |
| topic | EGFR de2-7EGFR extracellular domain mutation autoactivation dimerization disulfide bond free cysteine |
| url | http://www.mdpi.com/2072-6694/3/2/2032/ |
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