Affibody-mediated retention of the epidermal growth factor receptor in the secretory compartments leads to inhibition of phosphorylation in the kinase domain

Vernet E, Lundberg E, Friedman M, Rigamonti N, Klausing S, Nygren P-A, Gräslund T (2009)
New Biotechnology 25(6): 417-423.

Journal Article | Published | English

No fulltext has been uploaded

Author
; ; ; ; ; ;
Abstract
Abnormal activity of the epidermal growth factor receptor (EGFR) is associated with various cancer-related processes and motivates the search for strategies that can selectively block EGFR signalling. In this study, functional knockdown of EGFR was achieved through expression of an affibody construct, (ZEGFR:1907)(2-)KDEL, with high affinity for EGFR and extended with the amino acids KDEL to make it resident in the secretory compartments. Expression of (ZEGFR:1907)(2-)KDEL resulted in 80% reduction ofthe cell surface level of EGFR, and fluorescent staining for EGFR and the (ZEGFR:1907)(2-)KDEL construct showed overlapping intracellular localisation. Immunocapture of EGFR from cell lysates showed that an intracellular complex between EGFR and the affibody construct had been formed, further indicating aspecific interaction between the affibody construct and EGFR. Surface depletion of EGFR led to a dramatic decrease in the amount of kinase domain phosphorylated EGFR, coincident with a significant decrease in the proliferation rate.
Publishing Year
ISSN
PUB-ID

Cite this

Vernet E, Lundberg E, Friedman M, et al. Affibody-mediated retention of the epidermal growth factor receptor in the secretory compartments leads to inhibition of phosphorylation in the kinase domain. New Biotechnology. 2009;25(6):417-423.
Vernet, E., Lundberg, E., Friedman, M., Rigamonti, N., Klausing, S., Nygren, P. - A., & Gräslund , T. (2009). Affibody-mediated retention of the epidermal growth factor receptor in the secretory compartments leads to inhibition of phosphorylation in the kinase domain. New Biotechnology, 25(6), 417-423.
Vernet, E., Lundberg, E., Friedman, M., Rigamonti, N., Klausing, S., Nygren, P. - A., and Gräslund , T. (2009). Affibody-mediated retention of the epidermal growth factor receptor in the secretory compartments leads to inhibition of phosphorylation in the kinase domain. New Biotechnology 25, 417-423.
Vernet, E., et al., 2009. Affibody-mediated retention of the epidermal growth factor receptor in the secretory compartments leads to inhibition of phosphorylation in the kinase domain. New Biotechnology, 25(6), p 417-423.
E. Vernet, et al., “Affibody-mediated retention of the epidermal growth factor receptor in the secretory compartments leads to inhibition of phosphorylation in the kinase domain”, New Biotechnology, vol. 25, 2009, pp. 417-423.
Vernet, E., Lundberg, E., Friedman, M., Rigamonti, N., Klausing, S., Nygren, P.-A., Gräslund , T.: Affibody-mediated retention of the epidermal growth factor receptor in the secretory compartments leads to inhibition of phosphorylation in the kinase domain. New Biotechnology. 25, 417-423 (2009).
Vernet, Erik, Lundberg, Emma, Friedman, Mikaela, Rigamonti, Nicolò, Klausing, Sandra, Nygren, Per-Ake, and Gräslund , Torbjörn. “Affibody-mediated retention of the epidermal growth factor receptor in the secretory compartments leads to inhibition of phosphorylation in the kinase domain”. New Biotechnology 25.6 (2009): 417-423.
This data publication is cited in the following publications:
This publication cites the following data publications:

2 Citations in Europe PMC

Data provided by Europe PubMed Central.

Inhibitory effects of H-Ras/Raf-1-binding affibody molecules on synovial cell function.
Shibasaki S, Karasaki M, Graslund T, Nygren PA, Sano H, Iwasaki T., AMB Express 4(1), 2014
PMID: 26267111
Novel antigen design for the generation of antibodies to G-protein-coupled receptors.
Larsson K, Hofstrom C, Lindskog C, Hansson M, Angelidou P, Hokfelt T, Uhlen M, Wernerus H, Graslund T, Hober S., J. Immunol. Methods 370(1-2), 2011
PMID: 21605562

38 References

Data provided by Europe PubMed Central.

Intracellular expression of a single-chain antibody directed to the EGFR leads to growth inhibition of tumor cells.
Jannot CB, Beerli RR, Mason S, Gullick WJ, Hynes NE., Oncogene 13(2), 1996
PMID: 8710366
Intradiabodies, bispecific, tetravalent antibodies for the simultaneous functional knockout of two cell surface receptors.
Jendreyko N, Popkov M, Beerli RR, Chung J, McGavern DB, Rader C, Barbas CF 3rd., J. Biol. Chem. 278(48), 2003
PMID: 12947084
Phenotypic knockout of VEGF-R2 and Tie-2 with an intradiabody reduces tumor growth and angiogenesis in vivo.
Jendreyko N, Popkov M, Rader C, Barbas CF 3rd., Proc. Natl. Acad. Sci. U.S.A. 102(23), 2005
PMID: 15928093
Neu differentiation factor activation of ErbB-3 and ErbB-4 is cell specific and displays a differential requirement for ErbB-2.
Beerli RR, Graus-Porta D, Woods-Cook K, Chen X, Yarden Y, Hynes NE., Mol. Cell. Biol. 15(12), 1995
PMID: 8524214
Role of epidermal growth factor and ErbB2 receptors in 3T3-L1 adipogenesis.
Harrington M, Pond-Tor S, Boney CM., Obesity (Silver Spring) 15(3), 2007
PMID: 17372305
The role of individual SH2 domains in mediating association of phospholipase C-gamma1 with the activated EGF receptor
Chattopadhyay, J. Biol.Chem. 274(), 1999
Requirement of Ras-GTP-Raf complexes for activation of Raf-1 by protein kinase C.
Marais R, Light Y, Mason C, Paterson H, Olson MF, Marshall CJ., Science 280(5360), 1998
PMID: 9525855
HER receptor signaling confers resistance to the insulin-like growth factor-I receptor inhibitor, BMS-536924.
Haluska P, Carboni JM, TenEyck C, Attar RM, Hou X, Yu C, Sagar M, Wong TW, Gottardis MM, Erlichman C., Mol. Cancer Ther. 7(9), 2008
PMID: 18765823
A fully human recombinant IgG-like bispecific antibody to both the epidermal growth factor receptor and the insulin-like growth factor receptor for enhanced antitumor activity.
Lu D, Zhang H, Koo H, Tonra J, Balderes P, Prewett M, Corcoran E, Mangalampalli V, Bassi R, Anselma D, Patel D, Kang X, Ludwig DL, Hicklin DJ, Bohlen P, Witte L, Zhu Z., J. Biol. Chem. 280(20), 2005
PMID: 15757893
Construction and characterization of affibody-Fc chimeras produced in Escherichia coli.
Ronnmark J, Hansson M, Nguyen T, Uhlen M, Robert A, Stahl S, Nygren PA., J. Immunol. Methods 261(1-2), 2002
PMID: 11861078

Export

0 Marked Publications

Open Data PUB

Web of Science

View record in Web of Science®

Sources

PMID: 19552886
PubMed | Europe PMC

Search this title in

Google Scholar