Chemical Rescue of DeltaF508-CFTR Mimics Genetic Repair in Cystic Fibrosis Bronchial Epithelial Cells

Overview

Journal Mol Cell Proteomics

Specialties Biochemistry
Cell Biology
Molecular Biology

Date 2008 Feb 21

PMID 18285607

Citations 35

Authors

Om V Singh

Harvey B Pollard

Pamela L Zeitlin

Affiliations

Soon will be listed here.

Abstract

In a previous study of sodium 4-phenylbutyrate (4-PBA)-responsive proteins in cystic fibrosis (CF) IB3-1 bronchial epithelial cells, we identified 85 differentially expressed high abundance proteins from whole cellular lysate (Singh, O. V., Vij, N., Mogayzel, P. J., Jr., Jozwik, C., Pollard, H. B., and Zeitlin, P. L. (2006) Pharmacoproteomics of 4-phenylbutyrate-treated IB3-1 cystic fibrosis bronchial epithelial cells. J. Proteome Res. 5, 562-571). In the present work we hypothesize that a subset of heat shock proteins that interact with cystic fibrosis transmembrane conductance regulator (CFTR) in common during chemical rescue and genetic repair will identify therapeutic networks for targeted intervention. Immunocomplexes were generated from total cellular lysates, and three subcellular fractions (endoplasmic reticulum (ER), cytosol, and plasma membrane) with anti-CFTR polyclonal antibody from CF (IB3-1), chemically rescued CF (4-PBA-treated IB3-1), and genetically repaired CF (IB3-1/S9 daughter cells repaired by gene transfer with adeno-associated virus-(wild type) CFTR). CFTR-interacting proteins were analyzed on two-dimensional gels and identified by mass spectrometry. A set of 16 proteins known to act in ER-associated degradation were regulated in common and functionally connected to the protein processing, protein folding, and inflammatory response. Some of these proteins were modulated exclusively in ER, cytosol, or plasma membrane. A subset of 4-PBA-modulated ER-associated degradation chaperones (GRP94, HSP84, GRP78, GRP75, and GRP58) was observed to associate with the immature B form of CFTR in ER. HSP70 and HSC70 interacted with the C band (mature form) of CFTR at the cell surface. We conclude that chemically rescued CFTR associates with a specific set of HSP70 family proteins that mark therapeutic interactions and can be useful to correct both ion transport and inflammatory phenotypes in CF subjects.

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References

MEACHAM G, Patterson C, Zhang W, Younger J, Cyr D . The Hsc70 co-chaperone CHIP targets immature CFTR for proteasomal degradation. Nat Cell Biol. 2001; 3(1):100-5. DOI: 10.1038/35050509. View

Gassler C, Wiederkehr T, Brehmer D, Bukau B, Mayer M . Bag-1M accelerates nucleotide release for human Hsc70 and Hsp70 and can act concentration-dependent as positive and negative cofactor. J Biol Chem. 2001; 276(35):32538-44. DOI: 10.1074/jbc.M105328200. View

Pollard H, Ji X, Jozwik C, Jacobowitz D . High abundance protein profiling of cystic fibrosis lung epithelial cells. Proteomics. 2005; 5(8):2210-26. DOI: 10.1002/pmic.200401120. View

Shin B, Wang H, Yim A, Le Naour F, Brichory F, Jang J . Global profiling of the cell surface proteome of cancer cells uncovers an abundance of proteins with chaperone function. J Biol Chem. 2002; 278(9):7607-16. DOI: 10.1074/jbc.M210455200. View

Zeitlin P, Lu L, Rhim J, Cutting G, Stetten G, Kieffer K . A cystic fibrosis bronchial epithelial cell line: immortalization by adeno-12-SV40 infection. Am J Respir Cell Mol Biol. 1991; 4(4):313-9. DOI: 10.1165/ajrcmb/4.4.313. View

MELNICK J, Dul J, Argon Y . Sequential interaction of the chaperones BiP and GRP94 with immunoglobulin chains in the endoplasmic reticulum. Nature. 1994; 370(6488):373-5. DOI: 10.1038/370373a0. View

Gabriel S, Clarke L, Boucher R, Stutts M . CFTR and outward rectifying chloride channels are distinct proteins with a regulatory relationship. Nature. 1993; 363(6426):263-8. DOI: 10.1038/363263a0. View

Asea A, Kraeft S, Stevenson M, Chen L, Finberg R, Koo G . HSP70 stimulates cytokine production through a CD14-dependant pathway, demonstrating its dual role as a chaperone and cytokine. Nat Med. 2000; 6(4):435-42. DOI: 10.1038/74697. View

Pollard H, Eidelman O, Jozwik C, Huang W, Srivastava M, Ji X . De novo biosynthetic profiling of high abundance proteins in cystic fibrosis lung epithelial cells. Mol Cell Proteomics. 2006; 5(9):1628-37. DOI: 10.1074/mcp.M600091-MCP200. View

10.

Johnston J, Ward C, Kopito R . Aggresomes: a cellular response to misfolded proteins. J Cell Biol. 1998; 143(7):1883-98. PMC: 2175217. DOI: 10.1083/jcb.143.7.1883. View

11.

Lisanti M, Le Bivic A, Sargiacomo M, Rodriguez-Boulan E . Steady-state distribution and biogenesis of endogenous Madin-Darby canine kidney glycoproteins: evidence for intracellular sorting and polarized cell surface delivery. J Cell Biol. 1989; 109(5):2117-27. PMC: 2115858. DOI: 10.1083/jcb.109.5.2117. View

12.

Wallin R, Lundqvist A, More S, von Bonin A, Kiessling R, Ljunggren H . Heat-shock proteins as activators of the innate immune system. Trends Immunol. 2002; 23(3):130-5. DOI: 10.1016/s1471-4906(01)02168-8. View

13.

de Graeff-Meeder E, Rijkers G, Kuis W, van der Zee R, van Eden W, Zegers B . Antibodies to human HSP60 in patients with juvenile chronic arthritis, diabetes mellitus, and cystic fibrosis. Pediatr Res. 1993; 34(4):424-8. DOI: 10.1203/00006450-199310000-00008. View

14.

Liu Y, Liu W, Song X, Zuo J . Effect of GRP75/mthsp70/PBP74/mortalin overexpression on intracellular ATP level, mitochondrial membrane potential and ROS accumulation following glucose deprivation in PC12 cells. Mol Cell Biochem. 2005; 268(1-2):45-51. DOI: 10.1007/s11010-005-2996-1. View

15.

Vabulas R, Ahmad-Nejad P, Ghose S, Kirschning C, Issels R, Wagner H . HSP70 as endogenous stimulus of the Toll/interleukin-1 receptor signal pathway. J Biol Chem. 2002; 277(17):15107-12. DOI: 10.1074/jbc.M111204200. View

16.

Siebenlist U, Franzoso G, Brown K . Structure, regulation and function of NF-kappa B. Annu Rev Cell Biol. 1994; 10:405-55. DOI: 10.1146/annurev.cb.10.110194.002201. View

17.

Park K, Gaynor R, Kwak Y . Heat shock protein 27 association with the I kappa B kinase complex regulates tumor necrosis factor alpha-induced NF-kappa B activation. J Biol Chem. 2003; 278(37):35272-8. DOI: 10.1074/jbc.M305095200. View

18.

Farinha C, Nogueira P, Mendes F, Penque D, Amaral M . The human DnaJ homologue (Hdj)-1/heat-shock protein (Hsp) 40 co-chaperone is required for the in vivo stabilization of the cystic fibrosis transmembrane conductance regulator by Hsp70. Biochem J. 2002; 366(Pt 3):797-806. PMC: 1222832. DOI: 10.1042/BJ20011717. View

19.

Ferrari D, SOLING H . The protein disulphide-isomerase family: unravelling a string of folds. Biochem J. 1999; 339 ( Pt 1):1-10. PMC: 1220120. View

20.

Singh O, Vij N, Mogayzel Jr P, Jozwik C, Pollard H, Zeitlin P . Pharmacoproteomics of 4-phenylbutyrate-treated IB3-1 cystic fibrosis bronchial epithelial cells. J Proteome Res. 2006; 5(3):562-71. DOI: 10.1021/pr050319o. View