Mapping of cystic fibrosis transmembrane conductance regulator membrane topology by glycosylation site insertion

Xiu-Bao D Chang, Yue Xian Hou, Timothy J. Jensen, John R. Riordan

Research output: Contribution to journalArticle

112 Citations (Scopus)

Abstract

Technical difficulties in obtaining three-dimensional structures of intrinsic membrane proteins continues to limit understanding of their function. However, considerable insight can be gained from their two- dimensional topological arrangement in the lipid bilayer. Efficient molecular genetic approaches are available to discern the topology of prokaryotic but not of eukaryotic membrane proteins. The absolute asymmetry of the sidedness of their N-glycosylation was employed here to develop such a method using the cystic fibrosis transmembrane conductance regulator (CFTR). Insertion by in vitro mutagenesis of N-glycosylation consensus sequences (NXS/T) in predicted cytoplasmic and extracytoplasmic loops between hydrophobic sequences capable of traversing the membrane established the membrane topology of CFTR. This provides the first experimental evaluation of the original topological model of CFTR based solely on hydropathy algorithms and a method which may be generally applicable for the in vivo evaluation of the topology of other mammalian membrane proteins.

Original languageEnglish (US)
Pages (from-to)18572-18575
Number of pages4
JournalJournal of Biological Chemistry
Volume269
Issue number28
StatePublished - Jul 15 1994

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Glycosylation
Cystic Fibrosis Transmembrane Conductance Regulator
Membrane Proteins
Topology
Membranes
Mutagenesis
Lipid bilayers
Consensus Sequence
Lipid Bilayers
Molecular Biology

ASJC Scopus subject areas

  • Biochemistry

Cite this

Mapping of cystic fibrosis transmembrane conductance regulator membrane topology by glycosylation site insertion. / Chang, Xiu-Bao D; Hou, Yue Xian; Jensen, Timothy J.; Riordan, John R.

In: Journal of Biological Chemistry, Vol. 269, No. 28, 15.07.1994, p. 18572-18575.

Research output: Contribution to journalArticle

Chang, Xiu-Bao D ; Hou, Yue Xian ; Jensen, Timothy J. ; Riordan, John R. / Mapping of cystic fibrosis transmembrane conductance regulator membrane topology by glycosylation site insertion. In: Journal of Biological Chemistry. 1994 ; Vol. 269, No. 28. pp. 18572-18575.
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