Analysis of conventional and unconventional trafficking of CFTR and other membrane proteins

Heon Yung Gee, Joo Young Kim, Min Goo Lee

Research output: Contribution to journalArticle

Abstract

The cystic fibrosis transmembrane conductance regulator (CFTR) is a polytopic transmembrane protein that functions as a cAMP-activated anion channel at the apical membrane of epithelial cells. Mutations in CFTR cause cystic fibrosis and are also associated with monosymptomatic diseases in the lung, pancreas, intestines, and vas deferens. Many disease-causing CFTR mutations, including the deletion of a single phenylalanine residue at position 508 (ΔF508-CFTR), result in protein misfolding and trafficking defects. Therefore, intracellular trafficking of wild-type and mutant CFTR has been studied extensively, and results from these studies significantly contribute to our general understanding of mechanisms involved in the cell-surface trafficking of membrane proteins. CFTR is a glycoprotein that undergoes complex N-glycosylation as it passes through Golgi-mediated conventional exocytosis. Interestingly, results from recent studies revealed that CFTR and other membrane proteins can reach the plasma membrane via an unconventional alternative route that bypasses Golgi in specific cellular conditions. Here, we describe methods that have been used to investigate the conventional and unconventional surface trafficking of CFTR. With appropriate modifications, the protocols described in this chapter can also be applied to studies investigating the intracellular trafficking of other plasma membrane proteins.

Original languageEnglish
Pages (from-to)137-154
Number of pages18
JournalMethods in Molecular Biology
Volume1270
DOIs
Publication statusPublished - 2015 Jan 1

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Cystic Fibrosis Transmembrane Conductance Regulator
Membrane Proteins
Cell Membrane
Vas Deferens
Sequence Deletion
Exocytosis
Protein Transport
Phenylalanine
Glycosylation
Cystic Fibrosis
Lung Diseases
Intestines
Anions
Blood Proteins
Pancreas
Glycoproteins
Epithelial Cells
Mutation
Membranes

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Cite this

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Analysis of conventional and unconventional trafficking of CFTR and other membrane proteins. / Gee, Heon Yung; Kim, Joo Young; Lee, Min Goo.

In: Methods in Molecular Biology, Vol. 1270, 01.01.2015, p. 137-154.

Research output: Contribution to journalArticle

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