A431 cell variants lacking the blood group A antigen display increased high affinity epidermal growth factor-receptor number, protein-tyrosine kinase activity, and receptor turnover

doi:10.1083/jcb.107.3.939

This Article

	Full Text (PDF, 3154K)
	Alert me when this article is cited
	Citation Map

Services

	Email this article
	Similar articles in this journal
	Similar articles in PubMed
	Alert me to new content in the JCB
	Download to citation manager

Citing Articles

	Citing Articles via HighWire
	Citing Articles via CrossRef
	Citing Articles via Google Scholar

Google Scholar

	Articles by Defize, L. H.
	Articles by de Laat, S. W.
	Search for Related Content

PubMed

	PubMed Citation
	Articles by Defize, L. H.
	Articles by de Laat, S. W.


Social Bookmarking

	What's this?

ARTICLES

A431 cell variants lacking the blood group A antigen display increased high affinity epidermal growth factor-receptor number, protein-tyrosine kinase activity, and receptor turnover

LH Defize, DJ Arndt-Jovin, TM Jovin, J Boonstra, J Meisenhelder, T Hunter, HT de Hey and SW de Laat
Hubrecht Laboratory, Netherlands Institute for Developmental Biology, Utrecht.

The epidermal growth factor receptor (EGF-R) of human A431 cells bears an antigenic determinant that is closely related to the human blood group A carbohydrate structure. Labeling studies with blood group A reactive anti-EGF-R monoclonal antibodies and various lectins revealed that A431 cultures are heterogeneous with respect to blood group A expression. We have isolated clonal variants of these cells that either express (A431A+ cells) or completely lack (A431A- cells) the blood group A specific N-acetyl-D-galactosamine (GalNAc) residue. We show that this difference is due to the absence of a UDP-GalNAc:Gal transferase activity in A431A- cells. Subsequently, we have compared EGF-R functioning in these cell lines. Scatchard analysis of EGF- binding shows that in A431A- cells 6.3% of the EGF-R belongs to a high affinity subclass (Kd = 0.4 nM) while in A431A+ this subclass represents only 3.2% of the total receptor pool. The elevated level of high affinity receptors in A431A- cells is accompanied by a parallel increase in receptor protein- tyrosine kinase activity. In membrane preparations of A431A- cells, receptor autophosphorylation as well as phosphorylation of a tyrosine-containing peptide substrate is 2-3-fold higher as compared with A431A+ cells. In intact A431A-cells, the difference in receptor activity is measured as a 2-3-fold elevated level of receptor phosphorylation and a 2-3-fold higher abundance of phosphotyrosine in total cellular protein in A431A- cells. In addition, [35S]methionine pulse-chase experiments showed a ligand-independent increase in turnover of EGF-R in A431A- cells: the receptor's half life in these cells is 10 h as compared with 17 h in A431A+ cells. Our results suggest a possible involvement of GalNAc residue(s) in determining EGF-R affinity, protein-tyrosine kinase activity and turnover in A431 cells. Furthermore, our results indicate that high affinity EGF-R are the biologically active species with respect to protein-tyrosine kinase activity.
Add to CiteULike CiteULike Add to Complore Complore Add to Connotea Connotea Add to Del.icio.us Del.icio.us Add to Digg Digg Facebook Add to Reddit Reddit Add to Technorati Technorati Twitter What's this?

This article has been cited by other articles:

Francoual, M., Etienne-Grimaldi, M.-C., Formento, J.-L., Benchimol, D., Bourgeon, A., Chazal, M., Letoublon, C., Andre, T., Gilly, N., Delpero, J.-R., Lasser, P., Spano, J.-P., Milano, G. (2006). EGFR in colorectal cancer: more than a simple receptor. Ann Oncol 17: 962-967 [Abstract] [Full Text]
Wang, X., Gu, J., Ihara, H., Miyoshi, E., Honke, K., Taniguchi, N. (2006). Core Fucosylation Regulates Epidermal Growth Factor Receptor-mediated Intracellular Signaling. J. Biol. Chem. 281: 2572-2577 [Abstract] [Full Text]
Chung, K. Y., Shia, J., Kemeny, N. E., Shah, M., Schwartz, G. K., Tse, A., Hamilton, A., Pan, D., Schrag, D., Schwartz, L., Klimstra, D. S., Fridman, D., Kelsen, D. P., Saltz, L. B. (2005). Cetuximab Shows Activity in Colorectal Cancer Patients With Tumors That Do Not Express the Epidermal Growth Factor Receptor by Immunohistochemistry. JCO 23: 1803-1810 [Abstract] [Full Text]
Dorscheid, D. R., Wojcik, K. R., Yule, K., White, S. R. (2001). Role of cell surface glycosylation in mediating repair of human airway epithelial cell monolayers. Am. J. Physiol. Lung Cell. Mol. Physiol. 281: L982-L992 [Abstract] [Full Text]
Stroop, C. J.M., Weber, W., Gerwig, G. J., Nimtz, M., Kamerling, J. P., Vliegenthart, J. F.G. (2000). Characterization of the carbohydrate chains of the secreted form of the human epidermal growth factor receptor. Glycobiology 10: 901-917 [Abstract] [Full Text]
Menoret, A, Otry, C, Labarriere, N, Breimer, M., Piller, F, Meflah, K, Le Pendu, J (1995). The expression of carbohydrate blood group antigens correlates with heat resistance. J. Cell Sci. 108: 1691-1701 [Abstract]
Purushotham, K.R., Nakagawa, Y., Humphreys-Beher, M.G., Maeda, N., Schneyer, C.A. (1993). Rat Parotid Gland Acinar Cell Proliferation: Signal Transduction at the Plasma Membrane. CROBM 4: 537-543 [Abstract] [Full Text]