Editors' Choice

You have free access to this content

Genetic analysis of G protein-coupled receptor expression in Escherichia coli: Inhibitory role of DnaJ on the membrane integration of the human central cannabinoid receptor

  1. Georgios Skretas1,2,
  2. George Georgiou1,2,3,4,*

Article first published online: 22 AUG 2008

DOI: 10.1002/bit.22097

Issue

Biotechnology and Bioengineering

Biotechnology and Bioengineering

Volume 102, Issue 2, pages 357–367, 1 February 2009

Additional Information

How to Cite

Skretas, G. and Georgiou, G. (2009), Genetic analysis of G protein-coupled receptor expression in Escherichia coli: Inhibitory role of DnaJ on the membrane integration of the human central cannabinoid receptor. Biotechnol. Bioeng., 102: 357–367. doi: 10.1002/bit.22097

Author Information

  1. 1

    Department of Chemical Engineering, University of Texas at Austin, 2500 Speedway, Austin, Texas 78712; telephone: 512-471-6975; fax: 512-471-7963

  2. 2

    Institute for Cellular and Molecular Biology, University of Texas at Austin, 2500 Speedway, Austin, Texas 78712

  3. 3

    Department of Biomedical Engineering, University of Texas at Austin, 2500 Speedway, Austin, Texas 78712

  4. 4

    Section of Microbiology and Molecular Genetics, University of Texas at Austin, 2500 Speedway, Austin, Texas 78712

*Department of Chemical Engineering, University of Texas at Austin, 2500 Speedway, Austin, Texas 78712; telephone: 512-471-6975; fax: 512-471-7963.

Publication History

  1. Issue published online: 17 DEC 2008
  2. Article first published online: 22 AUG 2008
  3. Accepted manuscript online: 22 AUG 2008 12:00AM EST
  4. Manuscript Accepted: 12 AUG 2008
  5. Manuscript Revised: 28 JUL 2008
  6. Manuscript Received: 10 JUN 2008

Funded by

  • Welch Foundation
  • National Institute of Health. Grant Number: NIH GM 55090

SEARCH

SEARCH BY CITATION

ARTICLE TOOLS

Get PDF (644K)

References

  • Alper H, Stephanopoulos G. 2007. Global transcription machinery engineering: A new approach for improving cellular phenotype. Metab Eng 9(3): 258–267.
  • Alper H, Miyaoku K, Stephanopoulos G. 2005. Construction of lycopene-overproducing E. coli strains by combining systematic and combinatorial gene knockout targets. Nat Biotechnol 23(5): 612–616.
  • Andersson H, D'Antona AM, Kendall DA, Von Heijne G, Chin CN. 2003. Membrane assembly of the cannabinoid receptor 1: Impact of a long N-terminal tail. Mol Pharmacol 64(3): 570–577.
  • Bane SE, Velasquez JE, Robinson AS. 2007. Expression and purification of milligram levels of inactive G-protein coupled receptors in E. coli. Protein Expr Purif 52(2): 348–355.
  • Bukau B, Walker GC. 1989. Cellular defects caused by deletion of the Escherichia coli dnaK gene indicate roles for heat shock protein in normal metabolism. J Bacteriol 171(5): 2337–2346.
  • Calandra B, Tucker J, Shire D, Grisshammer R. 1997. Expression in Escherichia coli and characterisation of the human central CB1 and peripheral CB2 cannabinoid receptors. Biotechnol Lett 19(5): 425–428.
  • Chen Y, Song J, Sui SF, Wang DN. 2003. DnaK and DnaJ facilitated the folding process and reduced inclusion body formation of magnesium transporter CorA overexpressed in Escherichia coli. Protein Expr Purif 32(2): 221–231.
  • Cherezov V, Rosenbaum DM, Hanson MA, Rasmussen SG, Thian FS, Kobilka TS, Choi HJ, Kuhn P, Weis WI, Kobilka BK, et al. 2007. High-resolution crystal structure of an engineered human beta2-adrenergic G protein-coupled receptor. Science 318(5854): 1258–1265.
  • Cormack BP, Valdivia RH, Falkow S. 1996. FACS-optimized mutants of the green fluorescent protein (GFP). Gene 173(1 Spec No): 33–38.
  • Datsenko KA, Wanner BL. 2000. One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc Natl Acad Sci USA 97(12): 6640–6645.
  • Di Marzo V, Bifulco M, De Petrocellis L. 2004. The endocannabinoid system and its therapeutic exploitation. Nat Rev Drug Discov 3(9): 771–784.
  • Drew DE, von Heijne G, Nordlund P, de Gier JW. 2001. Green fluorescent protein as an indicator to monitor membrane protein overexpression in Escherichia coli. FEBS Lett 507(2): 220–224.
  • Geertsma ER, Groeneveld M, Slotboom DJ, Poolman B. 2008. Quality control of overexpressed membrane proteins. Proc Natl Acad Sci USA 105(15): 5722–5727.
  • Grisshammer R, Duckworth R, Henderson R. 1993. Expression of a rat neurotensin receptor in Escherichia coli. Biochem J 295(Pt 2): 571–576.
  • Guzman LM, Belin D, Carson MJ, Beckwith J. 1995. Tight regulation, modulation, and high-level expression by vectors containing the arabinose PBAD promoter. J Bacteriol 177(14): 4121–4130.
  • Haldimann A, Fisher SL, Daniels LL, Walsh CT, Wanner BL. 1997. Transcriptional regulation of the Enterococcus faecium BM4147 vancomycin resistance gene cluster by the VanS-VanR two-component regulatory system in Escherichia coli K-12. J Bacteriol 179(18): 5903–5913.
  • Hampe W, Voss RH, Haase W, Boege F, Michel H, Reilander H. 2000. Engineering of a proteolytically stable human beta 2-adrenergic receptor/maltose-binding protein fusion and production of the chimeric protein in Escherichia coli and baculovirus-infected insect cells. J Biotechnol 77(2–3): 219–234.
  • Hendrick JP, Langer T, Davis TA, Hartl FU, Wiedmann M. 1993. Control of folding and membrane translocation by binding of the chaperone DnaJ to nascent polypeptides. Proc Natl Acad Sci USA 90(21): 10216–10220.
  • Huang HC, Sherman MY, Kandror O, Goldberg AL. 2001. The molecular chaperone DnaJ is required for the degradation of a soluble abnormal protein in Escherichia coli. J Biol Chem 276(6): 3920–3928.
  • Jacoby E, Bouhelal R, Gerspacher M, Seuwen K. 2006. The 7 TM G-protein-coupled receptor target family. ChemMedChem 1(8): 761–782.
    Direct Link:
  • Kang MJ, Lee YM, Yoon SH, Kim JH, Ock SW, Jung KH, Shin YC, Keasling JD, Kim SW. 2005. Identification of genes affecting lycopene accumulation in Escherichia coli using a shot-gun method. Biotechnol Bioeng 91(5): 636–642.
    Direct Link:
  • Kiefer H. 2003. In vitro folding of alpha-helical membrane proteins. Biochim Biophys Acta 1610(1): 57–62.
  • Kristiansen K. 2004. Molecular mechanisms of ligand binding, signaling, and regulation within the superfamily of G-protein-coupled receptors: Molecular modeling and mutagenesis approaches to receptor structure and function. Pharmacol Ther 103(1): 21–80.
  • Link AJ, Skretas G, Strauch EM, Chari NS, Georgiou G. 2008. Efficient production of membrane-integrated and detergent-soluble G protein-coupled receptors in Escherichia coli. Protein Sci Advanced online publication July 1. DOI: 10.1110/ps.035980.108
    Direct Link:
  • Loll PJ. 2003. Membrane protein structural biology: The high throughput challenge. J Struct Biol 142(1): 144–153.
  • Luirink J, von Heijne G, Houben E, de Gier JW. 2005. Biogenesis of inner membrane proteins in Escherichia coli. Annu Rev Microbiol 59: 329–355.
  • Mayer MP, Bukau B. 2005. Hsp70 chaperones: Cellular functions and molecular mechanism. Cell Mol Life Sci 62(6): 670–684.
  • McCusker EC, Bane SE, O'Malley MA, Robinson AS. 2007. Heterologous GPCR expression: A bottleneck to obtaining crystal structures. Biotechnol Prog 23(3): 540–547.
    Direct Link:
  • Miroux B, Walker JE. 1996. Over-production of proteins in Escherichia coli: Mutant hosts that allow synthesis of some membrane proteins and globular proteins at high levels. J Mol Biol 260(3): 289–298.
  • Molina DM, Cornvik T, Eshaghi S, Haeggstrom JZ, Nordlund P, Sabet MI. 2008. Engineering membrane protein overproduction in Escherichia coli. Protein Sci 17: 673–680.
    Direct Link:
  • Murakami M, Kouyama T. 2008. Crystal structure of squid rhodopsin. Nature 453(7193): 363–367.
  • Palczewski K, Kumasaka T, Hori T, Behnke CA, Motoshima H, Fox BA, Le Trong I, Teller DC, Okada T, Stenkamp RE, et al. 2000. Crystal structure of rhodopsin: A G protein-coupled receptor. Science 289 (5480): 739–745.
  • Qiu XB, Shao YM, Miao S, Wang L. 2006. The diversity of the DnaJ/Hsp40 family, the crucial partners for Hsp70 chaperones. Cell Mol Life Sci 63(22): 2560–2570.
  • Rahav-Manor O, Carmel O, Karpel R, Taglicht D, Glaser G, Schuldiner S, Padan E. 1992. NhaR, a protein homologous to a family of bacterial regulatory proteins (LysR), regulates nhaA, the sodium proton antiporter gene in Escherichia coli. J Biol Chem 267(15): 10433–10438.
  • Raine A, Ullers R, Pavlov M, Luirink J, Wikberg JE, Ehrenberg M. 2003. Targeting and insertion of heterologous membrane proteins in E. coli. Biochimie 85(7): 659–668.
  • Rasmussen SG, Choi HJ, Rosenbaum DM, Kobilka TS, Thian FS, Edwards PC, Burghammer M, Ratnala VR, Sanishvili R, Fischetti RF, et al. 2007. Crystal structure of the human beta2 adrenergic G-protein-coupled receptor. Nature 450(7168): 383–387.
  • Santini CL, Bernadac A, Zhang M, Chanal A, Ize B, Blanco C, Wu LF. 2001. Translocation of jellyfish green fluorescent protein via the Tat system of Escherichia coli and change of its periplasmic localization in response to osmotic up-shock. J Biol Chem 276(11): 8159–8164.
  • Sarramegna V, Talmont F, Demange P, Milon A. 2003. Heterologous expression of G-protein-coupled receptors: Comparison of expression systems from the standpoint of large-scale production and purification. Cell Mol Life Sci 60(8): 1529–1546.
  • Shire D, Calandra B, Delpech M, Dumont X, Kaghad M, Le Fur G, Caput D, Ferrara P. 1996. Structural features of the central cannabinoid CB1 receptor involved in the binding of the specific CB1 antagonist S. J Biol Chem 271(12): 6941–6946.
  • Skerra A. 1994. Use of the tetracycline promoter for the tightly regulated production of a murine antibody fragment in Escherichia coli. Gene 151(1–2): 131–135.
  • Tucker J, Grisshammer R. 1996. Purification of a rat neurotensin receptor expressed in Escherichia coli. Biochem J 317(Pt 3): 891–899.
  • Voloshin ON, Vanevski F, Khil PP, Camerini-Otero RD. 2003. Characterization of the DNA damage-inducible helicase DinG from Escherichia coli. J Biol Chem 278(30): 28284–28293.
  • Weiss HM, Grisshammer R. 2002. Purification and characterization of the human adenosine A(2a) receptor functionally expressed in Escherichia coli. Eur J Biochem 269(1): 82–92.
    Direct Link:
  • Wild J, Altman E, Yura T, Gross CA. 1992. DnaK and DnaJ heat shock proteins participate in protein export in Escherichia coli. Genes Dev 6(7): 1165–1172.
  • Wild J, Rossmeissl P, Walter WA, Gross CA. 1996. Involvement of the DnaK-DnaJ-GrpE chaperone team in protein secretion in Escherichia coli. J Bacteriol 178(12): 3608–3613.
  • Yalovsky S, Paulsen H, Michaeli D, Chitnis PR, Nechushtai R. 1992. Involvement of a chloroplast HSP70 heat shock protein in the integration of a protein (light-harvesting complex protein precursor) into the thylakoid membrane. Proc Natl Acad Sci USA 89(12): 5616–5619.
  • Yeliseev A, Zoubak L, Gawrisch K. 2007. Use of dual affinity tags for expression and purification of functional peripheral cannabinoid receptor. Protein Expr Purif 53(1): 153–163.
Get PDF (644K)