Improving recombinant eukaryotic membrane protein yields in Pichia pastoris: The importance of codon optimization and clone selection

Molecular Membrane Biology

Volumn 28, Issue 6, 2011, Pages 398-411

  Öberg, Fredrik ^a   Sjöhamn, Jennie ^a   Conner, Matthew T ^b   Bill, Roslyn M ^b   Hedfalk, Kristina ^a  

^a UNIVERSITY OF GOTHENBURG   (Sweden)

^b ASTON UNIVERSITY   (United Kingdom)

Author keywords

integral membrane proteins; P. pastoris; Recombinant protein production

Indexed keywords

AQUAPORIN 1; AQUAPORIN 4; AQUAPORIN 8; CHIMERIC PROTEIN; GLYCOPEPTIDE; LITHIUM CHLORIDE; MEMBRANE FUSION PROTEIN; MEMBRANE PROTEIN; OLIGOMER; RECOMBINANT PROTEIN; UNCLASSIFIED DRUG; ZEOCIN;

ARTICLE; CARBOXY TERMINAL SEQUENCE; CLONE; CODON; CODON USAGE; CONTROLLED STUDY; ELECTROPORATION; HYDROPHILICITY; NONHUMAN; PICHIA PASTORIS; PRIORITY JOURNAL; PROCESS OPTIMIZATION; PROTEIN SYNTHESIS; SCREENING; SITE DIRECTED MUTAGENESIS; WATER TRANSPORT; WILD TYPE;

AQUAPORINS; CODON; ELECTROPORATION; HUMANS; LITHIUM CHLORIDE; PICHIA; RECOMBINANT PROTEINS; TRANSFORMATION, GENETIC;

BACTERIA (MICROORGANISMS); ESCHERICHIA COLI; EUKARYOTA; PICHIA PASTORIS;

EID: 80052008858     PISSN: 09687688     EISSN: 14645203     Source Type: Journal    
DOI: 10.3109/09687688.2011.602219     Document Type: Article

References (31)

1. Bill RM, Henderson PJ, Iwata S, Kunji ER, Michel H, Neutze R, Newstead S, Poolman B, Tate CG, Vogel H. 2011. Overcoming barriers to membrane protein structure determination. Nature Biotechnol 29:335-340.
2. Bonander N, Hedfalk K, Larsson C, Mostad P, Chang C, Gustafsson L, Bill RM. 2005. Design of improved membrane protein production experiments: Quantitation of the host response. Protein Sci 14:1729-1740. (Pubitemid 40994143)
3. Cereghino GP, Cereghino JL, IlgenC,Cregg JM. 2002. Production of recombinant proteins in fermenter cultures of the yeast Pichia pastoris. Curr Opin Biotechnol 13:329-332. (Pubitemid 35254090)
4. Chen X, Wang Q, Ni F, Ma J. Structure of the full-length Shaker potassium channel Kv1.2 by normal-mode-based X-ray crys-tallographic refinement. Proc Natl Acad Sci USA 2010;107: 11352-11357.
5. Conner MT, Conner AC, Brown JE, Bill RM. 2010. Membrane trafficking of aquaporin 1 is mediated by protein kinase C via microtubules and regulated by tonicity. Biochemistry 49:821-823.
6. Cregg JM, Tolstorukov I, Kusari A, Sunga J, Madden K, Chappell T. 2009. Expression in the yeast Pichia pastoris. Methods Enzymol 463:169-189.
7. Fantoni A, Bill RM, Gustafsson L, Hedfalk K. 2007. Improved yields of full-length functional human FGF1 can be achieved using the methylotrophic yeast Pichia pastoris. Protein Expr Purif 52:31-39. (Pubitemid 46038762)
8. Foster W, Helm A, Turnbull I, Gulati H, Yang B, Verkman AS, Skach WR. 2000. Identification of sequence determinants that direct different intracellular folding pathways for aquaporin-1 and aquaporin-4. J Biol Chem 275:34157-34165.
9. Gene Art. Method and device for optimising a nucleotide sequence for the purpose of expression of a protein. http://www.geneart.com/.
10. Gustafsson C, Govindarajan S, Minshull J. 2004. Codon bias and heterologous protein expression. Trends Biotechnol 22:346-353. (Pubitemid 38887543)
11. Hedfalk K, Pettersson N, Öberg F, Hohmann S, Gordon E. 2008. Production, characterization and crystallization of the Plasmodium falciparum aquaporin. Protein Expr Purif59:69-78.
12. Hennig L, Schafer E. 1998. Protein purification with C-terminal fusion of maltose binding protein. Protein Expr Purif 14:367-370. (Pubitemid 29020370)
13. Invitrogen C. 2005. EasySelect Pichia Expression Kit, version G. Accessed from the website: http://www.invitrogen.com/content/sfs/manuals/easyselect-man. pdf
14. Karlsson M, Fotiadis D, Sjovall S, Johansson I, Hedfalk K, Engel A, Kjellbom P. 2003. Reconstitution of water channel function of an aquaporin overexpressed and purified from Pichia pastoris. FEBS Lett 537:68-72. (Pubitemid 36246678)
15. Kremers GJ, Goedhart J, Van Den Heuvel DJ, Gerritsen HC, Gadella TW JR. 2007. Improved green and blue fluorescent proteins for expression in bacteria and mammalian cells. Biochemistry 46:3775-3783. (Pubitemid 46493477)
16. Lundstrom K. 2006. Structural genomics: The ultimate approach for rational drug design. Mol Biotechnol 34:205-212. (Pubitemid 46030394)
17. Nyblom M, Öberg F, Lindkvist-Petersson K, Hallgren K, Findlay H, Wikstrom J, Karlsson A, Hansson O, Booth PJ, Bill RM, Neutze R, Hedfalk K. 2007. Exceptional overproduction of a functional human membrane protein. Protein Expr Purif 56:110-120. (Pubitemid 47505316)
18. Öberg F, Ekvall M, Nyblom M, Backmark A, Neutze R, Hedfalk K. 2009. Insight into factors directing high production of eukaryotic membrane proteins; production of 13 human AQPs in Pichia pastoris. Mol Membr Biol 26:215-227.
19. Perez-Martin J, Cases I, De Lorenzo V. 1997. Design of a solubilization pathway for recombinant polypeptides in vivo through processing of a bi-protein with a viral protease. Protein Eng 10:725-730. (Pubitemid 27332081)
20. Petrovskaya LE, Shulga AA, Bocharova OV, Ermolyuk YS, Kryukova EA, Chupin VV, Blommers MJ, Arseniev AS, Kirpichnikov MP. Expression of G-protein coupled receptors in Escherichia coli for structural studies. Biochemistry (Mosc) 75:881-891.
21. Reizer J, Reizer A, Milton H, Saier J. 1993. The MIP family of integral membrane channel proteins: Sequence comparisons, evolutionary relationships, reconstructed pathway of evolution, and proposed functional differentiation of the two repeated halves of the proteins. Crit Rev Biochem Molec Biol 28: 235-257. (Pubitemid 23160986)
22. Roosild TP, Greenwald J, Vega M, Castronovo S, Riek R, Choe S. 2005. NMR structure of Mistic, a membrane-integrating protein for membrane protein expression. Science 307:1317-1321. (Pubitemid 40299982)
23. Roosild TP, Vega M, Castronovo S, Choe S. 2006. Characterization of the family of Mistic homologues. BMC Struct Biol 6:10.
24. Sharp PM, Li WH. 1987. The codon Adaptation Index-a measure of directional synonymous codon usage bias, and its potential applications. Nucleic Acids Res 15:1281-1295.
25. Sorensen HP. 2010. Towards universal systems for recombinant gene expression. Microbial Cell Factories 9:27.
26. Tate CG. 2001. Overexpression of mammalian integral membrane proteins for structural studies. FEBS Lett 504:94-98. (Pubitemid 32787055)
27. Tornroth-Horsefield S, Wang Y, Hedfalk K, Johanson U, Karlsson M, Tajkhorshid E, Neutze R, Kjellbom P. 2006. Structural mechanism of plant aquaporin gating. Nature 439:688-694. (Pubitemid 43237610)
28. Wang A, Clapper J, Guderian JA, Foy TM, Fanger GR, Retter MW, Skeiky YA. 2003. A novel method for increasing the expression level of recombinant proteins. Protein Expr Purif 30:124-133. (Pubitemid 36815241)
29. Werten MW, Wisselink WH, Jansen-Van Den Bosch TJ, De Bruin EC, De Wolf FA. 2001. Secreted production of a custom-designed, highly hydrophilic gelatin in Pichia pastoris. Protein Eng 14:447-454. (Pubitemid 32762382)
30. White SH. 2009. Biophysical dissection of membrane proteins. Nature 459:344-346.
31. Viard P, Butcher AJ, Halet G, Davies A, Nurnberg B, Heblich F, Dolphin AC. 2004. PI3K promotes voltage-dependent calcium channel trafficking to the plasma membrane. Nat Neurosci 7:939-946. (Pubitemid 39129502)