Defects in CFTR (cystic fibrosis transmembrane conductance regulator) maturation are central to the pathogenesis of CF (cystic fibrosis). Palmitoylation serves as a key regulator of maturational processing in other integral membrane proteins, but has not been tested previously for functional effects on CFTR. In the present study, we used metabolic labelling to confirm that wild-type and F508del CFTR are palmitoylated, and show that blocking palmitoylation with the pharmacologic inhibitor 2-BP (2-bromopalmitate) decreases steady-state levels of both wild-type and low temperature-corrected F508del CFTR, disrupts post-ER (endoplasmic reticulum) maturation and reduces ion channel function at the cell surface. PATs (protein acyl transferases) comprise a family of 23 gene products that contain a DHHC motif and mediate palmitoylation. Recombinant expression of specific PATs led to increased levels of CFTR protein and enhanced palmitoylation as judged by Western blot and metabolic labelling. Specifically, we show that DHHC-7 (i) increases steady-state levels of wild-type and F508del CFTR band B, (ii) interacts preferentially with the band B glycoform, and (iii) augments radiolabelling by [3H]palmitic acid. Interestingly, immunofluorescence revealed that DHHC-7 also sequesters the F508del protein to a post-ER (Golgi) compartment. Our findings point to the importance of palmitoylation during wild-type and F508del CFTR trafficking.
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Research Article|
March 28 2014
S-palmitoylation regulates biogenesis of core glycosylated wild-type and F508del CFTR in a post-ER compartment
Michelle L. McClure;
Michelle L. McClure
*Gregory Fleming James Cystic Fibrosis Research Center, University of Alabama at Birmingham, Birmingham, AL 35294, U.S.A.
†Department of Genetics, University of Alabama at Birmingham, Birmingham, AL 35294, U.S.A.
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Hui Wen;
Hui Wen
*Gregory Fleming James Cystic Fibrosis Research Center, University of Alabama at Birmingham, Birmingham, AL 35294, U.S.A.
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James Fortenberry;
James Fortenberry
*Gregory Fleming James Cystic Fibrosis Research Center, University of Alabama at Birmingham, Birmingham, AL 35294, U.S.A.
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Jeong S. Hong;
Jeong S. Hong
*Gregory Fleming James Cystic Fibrosis Research Center, University of Alabama at Birmingham, Birmingham, AL 35294, U.S.A.
‡Department of Cell, Developmental and Integrative Biology, University of Alabama at Birmingham, Birmingham, AL 35294, U.S.A.
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Eric J. Sorscher
Eric J. Sorscher
1
*Gregory Fleming James Cystic Fibrosis Research Center, University of Alabama at Birmingham, Birmingham, AL 35294, U.S.A.
§Department of Medicine, University of Alabama at Birmingham, Birmingham, AL 35294, U.S.A.
1To whom correspondence should be addressed (email sorscher@uab.edu).
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Publisher: Portland Press Ltd
Received:
August 02 2013
Revision Received:
January 15 2014
Accepted:
January 29 2014
Accepted Manuscript online:
January 29 2014
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2014 Biochemical Society
2014
Biochem J (2014) 459 (2): 417–425.
Article history
Received:
August 02 2013
Revision Received:
January 15 2014
Accepted:
January 29 2014
Accepted Manuscript online:
January 29 2014
Citation
Michelle L. McClure, Hui Wen, James Fortenberry, Jeong S. Hong, Eric J. Sorscher; S-palmitoylation regulates biogenesis of core glycosylated wild-type and F508del CFTR in a post-ER compartment. Biochem J 15 April 2014; 459 (2): 417–425. doi: https://doi.org/10.1042/BJ20131037
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