PMID: 9422381Jan 9, 1998Paper

Contributions of N-linked glycosylation to the expression of a functional alpha7-nicotinic receptor in Xenopus oocytes

Journal of Neurochemistry
D ChenJ W Patrick

Abstract

The alpha7 subunit of the neuronal nicotinic acetylcholine receptor, when expressed in Xenopus oocytes, forms homooligomeric ligand-gated ion channels that are blocked by a snake toxin, alpha-bungarotoxin. The amino-terminal extracellular domain of the alpha7 sequence has three consensus sites for asparagine-linked glycosylation (N46DS, N90MS, and N133AS). In this study, we show that alpha7 expressed either in vivo or in vitro is a glycoprotein of 57 kDa. In addition, we demonstrate by site-directed mutagenesis that all three consensus sites are used for glycosylation. To elucidate the role(s) of asparagine-linked glycosylation in the formation and function of the alpha7 receptor, wild-type and glycosylation-deficient alpha7 subunits were expressed in COS cells and oocytes. We examined biochemical and physiological properties of expressed receptors and found that alpha7 glycosylation mutations do not affect homooligomerization and surface protein expression of the alpha7 receptor but do affect surface expression of alpha-bungarotoxin binding sites and the function of the receptor. Our data indicate that asparagine-linked glycosylation is required for the expression of a functional alpha7 receptor in oocytes.

Citations

Mar 20, 2014·Glycobiology·Hilary Scott, Vladislav M Panin
Dec 15, 2006·Archives of Insect Biochemistry and Physiology·Jian-Rong GaoJeffrey G Scott
Mar 17, 2005·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·D B SattelleP C Biggin
May 2, 2000·The Journal of Biological Chemistry·K T Dineley, J W Patrick
Jul 20, 2010·The Journal of Biological Chemistry·Wen-Yi LoRobert L Macdonald
May 1, 2008·Molecular Membrane Biology·Neil S Millar, Patricia C Harkness
Dec 22, 2000·Journal of Molecular Biology·J P RogersR C Stevens
Aug 18, 2009·Neuroscience Research·John G Mielke, Geoffrey A R Mealing
Jun 15, 2011·Protein Expression and Purification·Shenping LiuEric T Baldwin

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