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Aptamers selected against the unglycosylated EGFRvIII ectodomain and delivered intracellularly reduce membrane-bound EGFRvIII and induce apoptosis.

Publication ,  Journal Article
Liu, Y; Kuan, C-T; Mi, J; Zhang, X; Clary, BM; Bigner, DD; Sullenger, BA
Published in: Biol Chem
February 2009

Epidermal growth factor receptor variant III (EGFRvIII) is a glycoprotein uniquely expressed in glioblastoma, but not in normal brain tissues. To develop targeted therapies for brain tumors, we selected RNA aptamers against the histidine-tagged EGFRvIII ectodomain, using an Escherichia coli system for protein expression and purification. Representative aptamer E21 has a dissociation constant (Kd) of 33x10(-9) m, and exhibits high affinity and specificity for EGFRvIII in ELISA and surface plasmon resonance assays. However, selected aptamers cannot bind the same protein expressed from eukaryotic cells because glycosylation, a post-translational modification present only in eukaryotic systems, significantly alters the structure of the target protein. By transfecting EGFRvIII aptamers into cells, we find that membrane-bound, glycosylated EGFRvIII is reduced and the percentage of cells undergoing apoptosis is increased. We postulate that transfected aptamers can interact with newly synthesized EGFRvIII, disrupt proper glycosylation, and reduce the amount of mature EGFRvIII reaching the cell surface. Our work establishes the feasibility of disrupting protein post-translational modifications in situ with aptamers. This finding is useful for elucidating the function of proteins of interest with various modifications, as well as dissecting signal transduction pathways.

Duke Scholars

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Published In

Biol Chem

DOI

ISSN

1431-6730

Publication Date

February 2009

Volume

390

Issue

2

Start / End Page

137 / 144

Location

Germany

Related Subject Headings

  • SELEX Aptamer Technique
  • RNA Processing, Post-Transcriptional
  • Molecular Sequence Data
  • Mice
  • ErbB Receptors
  • Enzyme-Linked Immunosorbent Assay
  • Cell Survival
  • Cell Membrane
  • Cell Line
  • Blotting, Western
 

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Liu, Y., Kuan, C.-T., Mi, J., Zhang, X., Clary, B. M., Bigner, D. D., & Sullenger, B. A. (2009). Aptamers selected against the unglycosylated EGFRvIII ectodomain and delivered intracellularly reduce membrane-bound EGFRvIII and induce apoptosis. Biol Chem, 390(2), 137–144. https://doi.org/10.1515/BC.2009.022
Liu, Yingmiao, Chien-Tsun Kuan, Jing Mi, Xiuwu Zhang, Bryan M. Clary, Darell D. Bigner, and Bruce A. Sullenger. “Aptamers selected against the unglycosylated EGFRvIII ectodomain and delivered intracellularly reduce membrane-bound EGFRvIII and induce apoptosis.Biol Chem 390, no. 2 (February 2009): 137–44. https://doi.org/10.1515/BC.2009.022.
Liu, Yingmiao, et al. “Aptamers selected against the unglycosylated EGFRvIII ectodomain and delivered intracellularly reduce membrane-bound EGFRvIII and induce apoptosis.Biol Chem, vol. 390, no. 2, Feb. 2009, pp. 137–44. Pubmed, doi:10.1515/BC.2009.022.
Liu Y, Kuan C-T, Mi J, Zhang X, Clary BM, Bigner DD, Sullenger BA. Aptamers selected against the unglycosylated EGFRvIII ectodomain and delivered intracellularly reduce membrane-bound EGFRvIII and induce apoptosis. Biol Chem. 2009 Feb;390(2):137–144.
Journal cover image

Published In

Biol Chem

DOI

ISSN

1431-6730

Publication Date

February 2009

Volume

390

Issue

2

Start / End Page

137 / 144

Location

Germany

Related Subject Headings

  • SELEX Aptamer Technique
  • RNA Processing, Post-Transcriptional
  • Molecular Sequence Data
  • Mice
  • ErbB Receptors
  • Enzyme-Linked Immunosorbent Assay
  • Cell Survival
  • Cell Membrane
  • Cell Line
  • Blotting, Western