Heterologous expression of G-protein-coupled receptors: Comparison of expression systems from the standpoint of large-scale production and purification

Cellular and Molecular Life Sciences

Volumn 60, Issue 8, 2003, Pages 1529-1546

  Sarramegna, V ^a   Talmont, F ^a   Demange, P ^a   Milon, A ^a  

^a INSTITUT DE PHARMACOLOGIE ET DE BIOLOGIE STRUCTURALE   (France)

Author keywords

GPCRs; Heterologous expression; Membrane proteins; NMR; Pichia pastoris; Receptor

Indexed keywords

G PROTEIN COUPLED RECEPTOR; MEMBRANE PROTEIN;

AMINO TERMINAL SEQUENCE; CARBOXY TERMINAL SEQUENCE; CELL MEMBRANE; CELL TYPE; ESCHERICHIA COLI; INSECT CELL; MAMMAL CELL; PROTEIN EXPRESSION; PROTEIN PROCESSING; PROTEIN PURIFICATION; REVIEW; SEQUENCE ANALYSIS; SIGNAL TRANSDUCTION; YEAST;

ANIMALS; BACULOVIRIDAE; CELL LINE; ESCHERICHIA COLI; GENE EXPRESSION; GTP-BINDING PROTEINS; HUMANS; MAMMALS; PICHIA; RECEPTORS, CELL SURFACE; RECOMBINANT PROTEINS; SACCHAROMYCES CEREVISIAE; SCHIZOSACCHAROMYCES; SPODOPTERA;

ESCHERICHIA COLI; INSECTA; MAMMALIA; PICHIA; PICHIA PASTORIS;

EID: 0041876269     PISSN: 1420682X     EISSN: None     Source Type: Journal    
DOI: 10.1007/s00018-003-3168-7     Document Type: Review

References (208)

1. Flower D. R. (1999) Modelling G-protein-coupled receptors for drug design. Biochim. Biophys. Acta 1422: 207-234
2. Bockaert J. and Pin J. P. (1998) [Use of a G-protein-coupled receptor to communicate. An evolutionary success]. C. R. Acad. Sci. III 321: 529-551
3. Bockaert J. and Pin J. P. (1999) Molecular tinkering of G protein-coupled receptors: an evolutionary success. EMBO J. 18: 1723-1729
4. Palczewski K., Kumasaka T., Hori T., Behnke C. A., Motoshima H., Fox B. A. et al. (2000) Crystal structure of rhodopsin: a G protein-coupled receptor. Science 289: 739-745
5. Schertler G. F. (1992) Overproduction of membrane proteins. Curr. Opin. Struct. Biol. 2: 534-544
6. Grisshammer R. and Tate C. G. (1995) Overexpression of integral membrane proteins for structural studies. Q. Rev. Biophys. 28: 315-422
7. King K, Dohlman H. G., Thorner J., Caron M. G. and Lefkowitz R. J. (1990) Control of yeast mating signal transduction by a mammalian beta 2-adrenergic receptor and Gs alpha subunit. Science 250: 121-123
8. Laage R. and Langosch D. (2001) Strategies for prokaryotic expression of eukaryotic membrane proteins. Traffic 2: 99-104
9. Waeber U., Buhr A., Schunk T. and Erni B. ( 1993) The glucose transporter of Escherichia coli. Purification and characterization by Ni+ chelate affinity chromatography of the IIBCGlc subunit. FEBS Lett. 324: 109-112
10. Sautel M. and Milligan G. (2000) Molecular manipulation of G-protein-coupled receptors: a new avenue into drug discovery. Curr. Med. Chem. 7: 889-896
11. Stanasila L., Pattus F. and Massotte D. (1998) Heterologous expression of G-protein-coupled receptors: human opioid receptors under scrutiny. Biochimie 80: 563-571
12. Wagner C. A., Friedrich B., Setiawan I., Lang F. and Broer S. (2000) The use of Xenopus laevis oocytes for the functional characterization of heterologously expressed membrane proteins. Cell. Physiol. Biochem. 10: 1-12
13. Tate C. G. and Grisshammer R. (1996) Heterologous expression of G-protein-coupled receptors. Trends Biotechnol. 14: 426-430
14. Grisshammer R., Duckworth R. and Henderson R. (1993) Expression of a rat neurotensin receptor in Escherichia coli. Biochem. J. 295: 571-576
15. Tucker J. and Grisshammer R. (1996) Purification of a rat neurotensin receptor expressed in Escherichia coli. Biochem. J. 317: 891-899
16. Stanasila L., Lim W. K., Neubig R. R. and Pattus F. (2000) Coupling efficacy and selectivity of the human mu-opioid receptor expressed as receptor-Galpha fusion proteins in Escherichia coli. J. Neurochem. 75: 1190-1199
17. Bertin B., Freissmuth M., Breyer R. M., Schutz W., Strosberg A. D. and Marullo S. (1992) Functional expression of the human serotonin 5-HT1A receptor in Escherichia coli. Ligand binding properties and interaction with recombinant G protein alpha-subunits. J. Biol. Chem. 267: 8200-8206
18. Kaushal S., Ridge K. D. and Khorana H. G. (1994) Structure and function in rhodopsin: the role of asparagine-linked glycosylation. Proc. Natl. Acad. Sci. USA 91: 4024-4028
19. Gibson S. K., Parkes J. H. and Liebman P. A. (1998) Phosphorylation stabilizes the active conformation of rhodopsin. Biochemistry 37: 13910
20. Gimpl G., Klein U., Reilander H. and Fahrenholz F. (1995) Expression of the human oxytocin receptor in baculovirus-infected insect cells: high-affinity binding is induced by a cholesterol-cyclodextrin complex. Biochemistry 34: 13794-1380
21. Nunez M. T. and Glass J. (1982) Reconstitution of the transferrin receptor in lipid vesicles. Effect of cholesterol on the binding of transferrin. Biochemistry 21: 4139-4143
22. Hasegawa J., Loh H. H. and Lee N. M. (1987) Lipid requirement for mu opioid receptor binding. J. Neurochem 49: 1007-1012
23. Lagane B., Gaibelet G., Meilhoc E., Masson J. M., Cezanne L. and Lopez A. (2000) Role of sterols in modulating the human micro-opioid receptor function in Saccharomyces cerevisiae. J. Biol. Chem. 275: 33197-33200
24. Abrahmsen L., Moks T., Nilsson B. and Uhlen M. (1986) Secretion of heterologous gene products to the culture medium of Escherichia coli. Nucleic. Acids. Res. 14: 7487-7500
25. Di Guan C., Li P., Riggs P. D. and Inouye H. (1988) Vectors that facilitate the expression and purification of foreign peptides in Escherichia coli by fusion to maltose-binding protein. Gene 67: 21-30
26. Kiefer H., Krieger J., Olszewski J. D., Von Heijne G., Prestwich G. D. and Breer H. (1996) Expression of an olfactory receptor in Escherichia coli: purification, reconstitution and ligand binding. Biochemistry 35: 16077-16084
27. LaVallie E. R. and McCoy J. M. (1995) Gene fusion expression systems in Escherichia coli. Curr. Opin. Biotechnol. 6: 501-506
28. Kiefer H., Maier K. and Vogel R. (1999) Refolding of G-protein-coupled receptors from inclusion bodies produced in Escherichia coli. Biochem. Soc. Trans. 27: 908-912
29. Efimov V. A., Fradkov A. F., Raskind A. B., Khristin M. S., Klimov V. V. and Chakhmakhcheva O. G. (1994) Expression of the barley psbA gene in Escherichia coli yields a functional in vitro photosystem II protein D1. FEBS Lett. 348: 153-157
30. Fiermonte G., Walker J. E. and Palmieri E (1993) Abundant bacterial expression and reconstitution of an intrinsic membrane-transport protein from bovine mitochondria. Biochem. J. 294: 293-299
31. Matsuyama S., Fujita Y. and Mizushima S. (1992) Large-scale production of membrane proteins fused to a truncated SecA in Escherichia coli. Biosci. Biotechnol. Biochem. 56: 1512-1514
32. Marullo S., Delavier-Klutchko C., Eshdat Y., Strosberg A. D. and Emorine L. (1988) Human beta 2-adrenergic receptors expressed in Escherichia coli membranes retain their pharmacological properties. Proc. Natl. Acad. Sci. USA 85: 7551-7555
33. Hampe W., Voss R. H., Haase W., Boege F., Michel H. and 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: 219-234
34. Chapot M. P., Eshdat Y., Marullo S., Guillet J. G., Charbit A., Strosberg A. D. et al. (1990) Localization and characterization of three different beta-adrenergic receptors expressed in Escherichia coli. Eur. J. Biochem. 187: 137-144
35. Emorine L. J., Marullo S., Briend-Sutren M. M., Patey G., Tate K., Delavier-Klutchko C. et al. (1989) Molecular characterization of the human beta 3-adrenergic receptor. Science 245: 1118-1121
36. Breyer R. M., Strosberg A. D. and Guillet J. G. (1990) Mutational analysis of ligand binding activity of beta 2 adrenergic receptor expressed in Escherichia coli. EMBO J. 9: 2679-2684
37. Grisshammer R., Little J. and Aharony D. (1994) Expression of rat NK-2 (neurokinin A) receptor in E. coli. Receptors Channels 2: 295-302
38. Blumer K. J., Reneke J. E. and Thorner J. (1988) The STE2 gene product is the ligand-binding component of the alpha-factor receptor of Saccharomyces cerevisiae. J. Biol. Chem. 263: 10836-10842
39. Kraakman L., Lemaire K., Ma P., Teunissen A. W., Donaton M. C., Van Dijck P. et al. (1999) A Saccharomyces cerevisiae G-protein coupled receptor, Gpr1, is specifically required for glucose activation of the cAMP pathway during the transition to growth on glucose. Mol. Microbiol. 32: 1002-1012
40. Price L. A., Kajkowski E. M., Hadcock J. R., Ozenberger B. A. and Pausch M. H. (1995) Functional coupling of a mammalian somatostatin receptor to the yeast pheromone response pathway. Mol. Cell. Biol. 15: 6188-6195
41. Stefan C. J., Overton M. C. and Blumer K. J. (1998) Mechanisms governing the activation and trafficking of yeast G protein- coupled receptors. Mol. Biol. Cell. 9: 885-899
42. Eckart M. R. and Bussineau C. M. (1996) Quality and authenticity of heterologous proteins synthesized in yeast. Curr. Opin. Biotechnol. 7: 525-530
43. Reilander H. and Weiss H. M. (1998) Production of G-protein-coupled receptors in yeast. Curr. Opin. Biotechnol. 9: 510-517
44. Payette P., Gossard F., Whiteway M. and Dennis M. (1990) Expression and pharmacological characterization of the human M1 muscarinic receptor in Saccharomyces cerevisiae. FEBS Lett. 266: 21-25
45. Sander P., Grunewald S., Bach M., Haase W, Reilander H. and Michel H. (1994) Heterologous expression of the human D2S dopamine receptor in protease- deficient Saccharomyces cerevisiae strains. Eur. J. Biochem 226: 697-705
46. Sander P., Grunewald S., Maul G., Reilander H. and Michel H. (1994) Constitutive expression of the human D2S-dopamine receptor in the unicellular yeast Saccharomyces cerevisiae. Biochim. Biophys. Acta 1193: 255-262
47. Sizmann D., Kuusinen H., Keranen S., Lomasney J., Caron M. G., Lefkowitz R. J. et al. (1996) Production of adrenergic receptors in yeast. Receptors Channels 4: 197-203
48. Hildebrandt V. (1989) Regeneration and functional incorporation of bacteriorhodopsin in membranes of fission yeast but not in E. coli. J. Protein Chem. 8: 345-346
49. Hildebrandt M., Saur U. and Nellen W. (1991) Structure, expression and inactivation by gene disruption of the Dictyostelium discoideum prespore gene EB4. Dev. Genet. 12: 163-169
50. Hildebrandt V., Fendler K., Heberle J., Hoffmann A., Bamberg E. and Buldt G. (1993) Bacteriorhodopsin expressed in Schizosaccharomyces pombe pumps protons through the plasma membrane. Proc. Natl. Acad. Sci. USA 90: 3578-3582
51. Maundrell K. (1993) Thiamine-repressible expression vectors pREP and pRIP for fission yeast. Gene 123: 127-130
52. Sander P., Grunewald S., Reilander H. and Michel H. (1994) Expression of the human D2S dopamine receptor in the yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe: a comparative study. FEBS Lett. 344: 41-46
53. Presland J. and Strange P. G. (1998) Pharmacological characterisation of the D2 dopamine receptor expressed in the yeast Schizosaccharomyces pombe. Biochem. Pharmacol. 56: 577-582
54. Arkinstall S., Edgerton M, Payton M. and Maundrell K. (1995) Co-expression of the neurokinin NK2 receptor and G-protein components in the fission yeast Schizosaccharomyces pombe. FEBS Lett. 375: 183-187
55. Ficca A. G., Testa L. and Tocchini Valentini G. P. (1995) The human beta 2-adrenergic receptor expressed in Schizosaccharomyces pombe retains its pharmacological properties. FEBS Lett. 377: 140-144
56. Cregg J. M., Cereghino J. L., Shi J. and Higgins D. R. (2000) Recombinant protein expression in Pichiapastoris. Mol. Biotechnol. 16: 23-52
57. Faber K. N., Harder W., Ab G. and Veenhuis M. (1995) Review: methylotrophic yeasts as factories for the production of foreign proteins. Yeast 11: 1331-1344
58. Gellissen G. (2000) Heterologous protein production in methylotrophic yeasts. Appl. Microbiol. Biotechnol. 54: 741-750
59. Higgins D. R. and Cregg J. M. (1998) Introduction to Pichia pastoris. Methods Mol. Biol. 103: 1-15
60. Hollenberg C. P. and Gellissen G. (1997) Production of recombinant proteins by methylotrophic yeasts. Curr. Opin. Biotechnol. 8: 554-560
61. Romanos M. A., Scorer C. A. and Clare J. J. (1992) Foreign gene expression in yeast: a review. Yeast 8: 423-488
62. Talmont F., Sidobre S., Demange P., Milon A. and Emorine L. J. (1996) Expression and pharmacological characterization of the human mu-opioid receptor in the methylotrophic yeast Pichiapastoris. FEBS Lett. 394: 268-272
63. Weiss H. M., Haase W., Michel H. and Reilander H. (1995) Expression of functional mouse 5-HT5A serotonin receptor in the methylotrophic yeast Pichia pastoris: pharmacological characterization and localization. FEBS Lett. 377: 451-456
64. Bach M., Sander P, Haase W. and Reilander H. (1996) Pharmacological and biochemical characterization of the mouse 5HT5A serotonin receptor heterologously produced in the yeast Saccharomyces cerevisiae. Receptors Channels 4: 129-139
65. Weiss H. M., Haase W. and Reilander H. (1998) Expression of an integral membrane protein, the 5HT5A receptor. Methods Mol. Biol. 103: 227-239
66. Weiss H. M., Haase W., Michel H. and Reilander H. (1998) Comparative biochemical and pharmacological characterization of the mouse 5HT5A 5-hydroxytryptamine receptor and the human beta2-adrenergic receptor produced in the methylotrophic yeast Pichia pastoris. Biochem. J. 330: 1137-1147
67. Gaibelet G., Meilhoc E., Riond J., Saves I., Exner T., Liaubet L. et al. (1999) Nonselective coupling of the human mu-opioid receptor to multiple inhibitory G-protein isoforms. Eur. J. Biochem. 261: 517-523
68. Sarramegna V., Demange P., Milon A. and Talmont F. (2002) Optimizing functional versus total expression of the human mu-opioid receptor in Pichia pastoris. Protein Expr. Purif. 24: 212-220
69. Sarramegna V., Talmont F., Milon A. and Demange P. (2002) Green fluorescent protein as a reporter of human μ-opioid receptor overexpresion in the methylotrophis yeast Pichia pastoris. J. Biotechnology 99: 23-39
70. Abdulaev N. G., Popp M. P., Smith W. C. and Ridge K. D. (1997) Functional expression of bovine opsin in the methylotrophic yeast Pichia pastoris. Protein Expr. Purif. 10: 61-69
71. Mollaaghababa R., Davidson F. F., Kaiser C. and Khorana H. G. (1996) Structure and function in rhodopsin: expression of functional mammalian opsin in Saccharomyces cerevisiae. Proc. Natl. Acad. Sci. USA 93: 11482-11486
72. David N. E., Gee M., Andersen B., Naider F., Thorner J. and Stevens R. C. (1997) Expression and purification of the Saccharomyces cerevisiae alpha-factor receptor (Ste2p), a 7-transmembrane-segment G protein-coupled receptor. J. Biol. Chem. 272: 15553-15561
73. Fraser M. J. (1992) The baculovirus-infected insect cell as a eukaryotic gene expression system. Curr. Top. Microbiol. Immunol. 158: 131-172
74. Pfeifer T. A. (1998) Expression of heterologous proteins in stable insect cell culture. Curr. Opin. Biotechnol. 9: 518-521.
75. Kempf J., Snook L. A., Vonesch J. L., Dahms T. E., Pattus F. and Massotte D. (2002) Expression of the human mu opioid receptor in a stable Sf9 cell line. J. Biotechnol. 95: 181-187
76. Bernard A. R., Kost T. A., Overton L., Cavegn C., Young J., Bertrand M. et al. (1994) Recombinant protein expression in a Drosophila cell line: comparison with the baculovirus system. Cytotechnology 15: 139-144
77. Kleymann G., Boege F., Hahn M., Hampe W., Vasudevan S. and Reilander H. (1993) Human beta 2-adrenergic receptor produced in stably transformed insect cells is functionally coupled via endogenous GTP-binding protein to adenylyl cyclase. Eur. J. Biochem 213: 797-804
78. Millar N. S., Baylis H. A., Reaper C., Bunting R., Mason W. T. and Sattelle D. B. (1995) Functional expression of a cloned Drosophila muscarinic acetylcholine receptor in a stable Drosophila cell line. J. Exp. Biol. 198: 1843-1850
79. Mulheron J. G., Casanas S. J., Arthur J. M., Gamovskaya M. N., Gettys T. W. and Raymond J. R. (1994) Human 5-HT1A receptor expressed in insect cells activates endogenous G(o)-like G protein(s). J. Biol. Chem. 269: 12954-12962
80. Mazina K. E., Strader C. D. and Fong T. M. (1994) Expression and solubilization of a recombinant human neurokinin-1 receptor in insect cells. J. Recept. Res. 14: 63-73
81. Rinken A., Kameyama K, Haga T. and Engstrom L. (1994) Solubilization of muscarinic receptor subtypes from baculovirus infected Sf9 insect cells. Biochem. Pharmacol. 48: 1245-1251
82. Guan X. M., Kobilka T. S. and Kobilka B. K. (1992) Enhancement of membrane insertion and function in a type IIIb membrane protein following introduction of a cleavable signal peptide. J. Biol. Chem 267: 21995-21998
83. Grunewald S., Reilander H. and Michel H. (1996) In vivo reconstitution of dopamine D2S receptor-mediated G protein activation in baculovirus-infected insect cells: preferred coupling to Gi1 versus Gi2. Biochemistry 35: 15162-15173
84. Reilander H., Boege F., Vasudevan S., Maul G., Hekman M, Dees C., et al. (1991) Purification and functional characterization of the human beta 2- adrenergic receptor produced in baculovirus-infected insect cells. FEBS Lett. 282: 441-444
85. Vasudevan S., Hulme E. C., Bach M., Haase W., Pavia J. and Reilander H. (1995) Characterization of the rat m3 muscarinic acetylcholine receptor produced in insect cells infected with recombinant baculovirus. Eur. J. Biochem. 227: 466-475
86. Harfst E., Johnstone A. P., Gout I., Taylor A. H., Waterfield M. D. and Nussey S. S. (1992)The use of the amplifiable high-expression vector pEE 14 to study the interactions of autoantibodies with recombinant human thyrotrophin receptor. Mol. Cell. Endocrinol. 83: 117-123
87. Boundy V. A., Lu L. and Molinoff P. B. (1996) Differential coupling of rat D2 dopamine receptor isoforms expressed in Spodoptera frugiperda insect cells. J. Pharmacol. Exp. Ther. 276: 784-794
88. Gimpl G. and Fahrenholz F. (2000) Human oxytocin receptors in cholesterol-rich vs. cholesterol-poor microdomains of the plasma membrane. Eur. J. Biochem 267: 2483-2497
89. Mouillac B., Caron M., Bonin H., Dennis M. and Bouvier M. (1992) Agonist-modulated palmitoylation of beta 2-adrenergic receptor in Sf9 cells. J. Biol. Chem. 267: 21733-21737
90. Nebigil C. G., Gamovskaya M. N., Casanas S. J., Mulheron J. G., Parker E. M., Gettys T. W. et al. (1995) Agonist-induced desensitization and phosphorylation of human 5-HT1A receptor expressed in Sf9 insect cells. Biochemistry 34: 11954-11962
91. Ng G. Y., O'Dowd B. F., Caron M., Dennis M., Brann M. R. and George S. R. (1994) Phosphorylation and palmitoylation of the human D2L dopamine receptor in Sf9 cells. J. Neurochem. 63: 1589-1595
92. Ausubel F. (1994). Current Protocols in Molecular Biology. Greene Publishing Associates and Wiley-Interscience, New York
93. Moss B. (1991) Vaccinia virus: a tool for research and vaccine development. Science 252: 1662-1667
94. Mackett M. (1994) Expression of membrane proteins using vaccinia recombinants. In: Membrane Protein Expression Systems: A User's Guide, pp. 125-175, Gould G. W. (ed.), Portland Press, London
95. Liljestrom P. and Garoff H. (1991) A new generation of animal cell expression vectors based on the Semliki Forest virus replicon. Biotechnology 9: 1356-1361
96. Schwartz T. W. (1994) Locating ligand-binding sites in 7TM receptors by protein engineering. Curr. Opin. Biotechnol. 5: 434-444
97. Strader C. D., long T. M., Tota M. R., Underwood D. and Dixon R. A. (1994) Structure and function of G protein-coupled receptors. Annu. Rev. Biochem. 63: 101-132
98. Lohse M. J. (1992) Stable overexpression of human beta 2-adrenergic receptors in mammalian cells. Naunyn Schmiedebergs Arch. Pharmacol. 345: 444-451
99. Reeves P. J., Kim J. M. and Khorana H. G. (2002) Structure and function in rhodopsin: a tetracycline-inducible system in stable mammalian cell lines for high-level expression of opsin mutants. Proc. Natl. Acad. Sci. USA 99: 13413-13418
100. Turner G. J., Reusch R., Winter-Vann A. M., Martinez L. and Betlach M. C. (1999) Heterologous gene expression in a membrane-protein-specific system. Protein Expr. Purif. 17: 312-323
101. Sohlemann P., Soppa J., Oesterhelt D. and Lohse M. J. (1997) Expression of beta 2-adrenoceptors in halobacteria. Naunyn Schmiedebergs Arch. Pharmacol. 355: 150-160
102. Pajot-Augy E., Couture L., Bozon V., Remy J. J., Biache G., Severini M. et al. (1995) High-level expression of recombinant porcine LH receptor in baculovirus-infected insect cells or caterpillars. J. Mol. Endocrinol. 14: 51-66
103. Eidne K. A. (1994) Expression of receptors in Xenopus oocytes. In: Membrane Protein Expression Systems: A User's Guide, pp. 275-300, Gould G.W. (led.), Portland Press, London
104. Eroglu C., Cronet P., Panneels V., Beaufils P. and Sinning I. (2002) Functional reconstitution of purified metabotropic glutamate receptor expressed in the fly eye. EMBO Rep. 3: 491-496
105. Alimov A. P., Khmelnitsky A., Simonenko P. N., Spirin A. S. and Chetverin A. B. (2000) Cell-free synthesis and affinity isolation of proteins on a nanomole scale. Biotechniques 28: 338-344
106. Baranov V. I. and Spirin A. S. (1993) Gene expression in cell-free system on preparative scale. Methods Enzymol. 217: 123-142
107. Kolb V. A., Makeyev E. V. and Spirin A. S. (2000) Co-translational folding of an eukaryotic multidomain protein in a prokaryotic translation system. J. Biol. Chem. 275: 16597-16601
108. Martemyanov K. A., Shirokov V. A., Kurnasov O. V., Gudkov A. T. and Spirin A. S. (2001) Cell-free production of biologically active polypeptides: application to the synthesis of antibacterial peptide cecropin. Protein Expr. Purif. 21: 456-461
109. Ryabova L. A., Morozov I. and Spirin A. S. (1998) Continuous-flow cell-free translation, transcription-translation and replication-translation systems. Methods Mol. Biol. 77: 179-193
110. Yokoyama S., Hirota H., Kigawa T., Yabuki T., Shirouzu M., Terada T. et al. (2000) Structural genomics projects in Japan. Nat. Struct. Biol. 7 Suppl: 943-945
111. Yokoyama S., Matsuo Y., Hirota H., Kigawa T., Shirouzu M., Kuroda Y. et al. (2000) Structural genomics projects in Japan. Prog. Biophys. Mol. Biol. 73: 363-376
112. Inooka H., Ohtaki T., Kitahara O., Ikegami T., Endo S., Kitada C. et al. (2001) Conformation of a peptide ligand bound to its G-protein coupled receptor. Nat. Struct. Biol. 8: 161-165
113. Marullo S., Delavier-Klutcho C., Guillet J. G., Charbit A., Strosberg A. D. and Emorine L. J. (1989) Expression of human betal and beta2 adrenergic receptors in E. coli as a new tool for ligand screening. Bio/Technology 7: 923-927
114. Hill R. A. and Sillence M. N. (1997) Improved membrane isolation in the purification of beta 2-adrenoceptors from transgenic Escherichia coli. Protein Expr. Purif. 10: 162-167
115. Lacatena R. M., Cellini A., Scavizzi F. and Tocchini-Valentini G. P. (1994) Topological analysis of the human beta 2-adrenergic receptor expressed in Escherichia coli. Proc. Natl. Acad. Sci. USA 91: 10521-10525
116. Freissmuth M., Selzer E., Marullo S., Schutz W. and Strosberg A. D. (1991) Expression of two human beta-adrenergic receptors in Escherichia coli: functional interaction with two forms of the stimulatory G protein. Proc. Natl. Acad. Sci. USA 88: 8548-8552
117. Haendler B, Hechler U., Becker A. and Schleuning W. D. (1993) Expression of human endothelin receptor ETB by Escherichia coli transformants. Biochem. Biophys. Res. Commun. 191: 633-638
118. Hanninen A. L., Bamford D. H. and Grisshammer R. (1994) Expression in Escherichia coli of rat neurotensin receptor fused to membrane proteins from the membrane-containing bacteriophage PRD1. Biol. Chem. Hoppe Seyler 375: 833-836
119. Jockers R., Linder M. E., Hohenegger M., Nanoff C., Bertin B., Strosberg A. D. et al. (1994) Species difference in the G protein selectivity of the human and bovine A1-adenosine receptor. J. Biol. Chem. 269: 32077-32084
120. Munch G., Walker P., Shine J. and Herzog H. (1995) Ligand binding analysis of human neuropeptide Y 1 receptor mutants expressed in E. coli. Receptors Channels 3: 291-297
121. Herzog H., Munch G. and Shine J. (1994) Human neuropeptide Y1 receptor expressed in Escherichia coli retains its pharmacological properties. DNA Cell Biol. 13: 1221-1225
122. Weiss H. M. and Grisshammer R. (2002) Purification and characterization of the human adenosine A2a receptor functionally expressed in Escherichia coli. Eur. J. Biochem. 269: 82-92
123. Schulein R., Rutz C. and Rosenthal W. (1996) Membrane targeting and determination of transmembrane topology of the human vasopressin V2 receptor. J. Biol. Chem. 271: 28844-28852
124. Stanasila L., Massotte D., Kieffer B. L. and Pattus F. (1999) Expression of delta, kappa and mu human opioid receptors in Escherichia coli and reconstitution of the high-affinity state for agonist with heterotrimeric G proteins. Eur. J. Biochem 260: 430-438
125. Furukawa H. and Haga T. (2000) Expression of functional M2 muscarinic acetylcholine receptor in Escherichia coli. J. Biochem. (Tokyo) 127: 151-161
126. Busuttil B. E., Turney K. L. and Frauman A. G. (2001) The expression of soluble, full-length, recombinant human TSH receptor in a prokaryotic system. Protein Expr. Purif. 23: 369-373
127. Schiller H., Haase W., Molsberger E., Janssen P., Michel H. and Reilander H. (2000) The human ET(B) endothelin receptor heterologously produced in the methylotrophic yeast Pichia pastoris shows high-affinity binding and induction of stacked membranes. Receptors Channels 7: 93-107
128. Huang H. J., Liao C. F., Yang B. C. and Kuo T. T. (1992) Functional expression of rat M5 muscarinic acetylcholine receptor in yeast. Biochem. Biophys. Res. Commun. 182: 1180-1186
129. Andersen B. and Stevens R. C. (1998) The human D1A dopamine receptor: heterologous expression in Saccharomyces cerevisiae and purification of the functional receptor. Protein Expr. Purif. 13: 111-119
130. Kajkowski E. M., Price L. A., Pausch M. H., Young K. H. and Ozenberger B. A. (1997) Investigation of growth hormone releasing hormone receptor structure and activity using yeast expression technologies. J. Recept. Signal Transduct. Res. 17: 293-303
131. Erickson J. R., Wu J. J., Goddard J. G., Tigyi G., Kawanishi K., Tomei L. D. et al. (1998) Edg-2/Vzg-1 couples to the yeast pheromone response pathway selectively in response to lysophosphatidic acid. J. Biol. Chem. 273: 1506-1510
132. Cronan J. E. Jr (1990) Biotination of proteins in vivo. A post-translational modification to label, purify and study proteins. J. Biol. Chem. 265: 10327-10333
133. George S. T., Arbabian M. A., Ruoho A. E., Kiely J. and Malbon C. C. (1989) High-efficiency expression of mammalian beta-adrenergic receptors in baculovirus-infected insect cells. Biochem. Biophys. Res. Commun. 163: 1265-1269
134. Parker E. M., Kameyama K, Higashijima T. and Ross E. M. (1991) Reconstitutively active G protein-coupled receptors purified from baculovirus-infected insect cells. J. Biol. Chem. 266: 519-527
135. Chidiac P., Hebert T. E., Valiquette M., Dennis M. and Bouvier M. (1994) Inverse agonist activity of beta-adrenergic antagonists. Mol. Pharmacol. 45: 490-499
136. Kwatra M. M., Schwinn D. A., Schreurs J., Blank J. L., Kim C. M., Benovic J. L. et al. (1993) The substance P receptor, which couples to Gq/11, is a substrate of beta-adrenergic receptor kinase 1 and 2. J. Biol. Chem. 268: 9161-9164
137. Loisel T. P., Ansanay H., St-Onge S., Gay B., Boulanger P., Strosberg A. D., et al. (1997) Recovery of homogeneous and functional beta 2-adrenergic receptors from extracellular baculovirus particles. Nat. Biotechnol. 15: 1300-1304
138. Kobilka B. K. (1995) Amino and carboxyl terminal modifications to facilitate the production and purification of a G protein-coupled receptor. Anal. Biochem. 231: 269-271
139. Parker E. M. and Ross E. M. (1991) Truncation of the extended carboxyl-terminal domain increases the expression and regulatory activity of the avian beta-adrenergic receptor. J. Biol. Chem. 266: 9987-9996
140. Oker-Blom C., Jansson C., Karp M., Lindqvist C., Savola J. M., Vlak J., et al. (1993) Functional analysis of the human alpha 2C-C4 adrenergic receptor in insect cells expressed by a luciferase-based baculovirus vector. Biochim. Biophys. Acta 1176: 269-275
141. Ng G. Y., George S. R., Zastawny R. L., Caron M., Bouvier M., Dennis M. et al. (1993) Human serotonin1B receptor expression in Sf9 cells: phosphorylation, palmitoylation, and adenylyl cyclase inhibition. Biochemistry 32: 11727-11733
142. Parker E. M., Grisel D. A., Iben L. G., Nowak H. P., Mahle C. D., Yocca F. D. et al. (1994) Characterization of human 5-HT1 receptors expressed in Sf9 insect cells. Eur. J. Pharmacol. 268: 43-53
143. Butkerait P., Zheng Y., Hallak H., Graham T. E., Miller H. A., Burris K. D. et al. (1995) Expression of the human 5-hydroxy-tryptamine1A receptor in Sf9 cells. Reconstitution of a coupled phenotype by co-expression of mammalian G protein subunits. J. Biol. Chem. 270: 18691-18699
144. Ponimaskin E. G., Schmidt M. F., Heine M., Bickmeyer U. and Richter D. W. (2001) 5-Hydroxytryptamine 4(a) receptor expressed in Sf9 cells is palmitoylated in an agonist-dependent manner. Biochem. J. 353: 627-634
145. Labrecque J., Fargin A., Bouvier M., Chidiac P. and Dennis M. (1995) Serotonergic antagonists differentially inhibit spontaneous activity and decrease ligand binding capacity of the rat 5-hydroxytryptamine type 2C receptor in Sf9 cells. Mol. Pharmacol. 48: 150-159
146. Aharony D., Little J. et al. (1994) "Isolation and characterization of neurokinin. A receptor cDNAs from guinea-pig lung and rabbit pulmonary artery." J Recept Res 14(6-8): 399-421
147. Mazina K. E., Strader C. D., Tota M. R., Daniel S. and Fong T. M. (1996) Purification and reconstitution of a recombinant human neurokinin-1 receptor. J. Recept. Signal Transduct. Res. 16: 191-207
148. Wehmeyer A. and Schulz R. (1997) Overexpression of delta-opioid receptors in recombinant baculovirus-infected Trichoplusia in 'High 5' insect cells. J. Neurochem 68: 1361-1371
149. Obermeier H., Wehmeyer A. and Schulz R. (1996) Expression of mu-, delta- and kappa-opioid receptors in baculovirus-infected insect cells. Eur. J. Pharmacol. 318: 161-166
150. Massotte D., Baroche L., Simonin F., Yu L., Kieffer B. and Pattus E (1997) Characterization of delta, kappa, and mu human opioid receptors overexpressed in baculovirus-infected insect cells. J. Biol. Chem. 272: 19987-19992
151. Massotte D., Pereira C. A., Pouliquen Y. and Pattus F. (1999) Parameters influencing human mu opioid receptor over-expression in baculovirus-infected insect cells. J. Biotechnol. 69: 39-45
152. Wei Q., Zhou D. H., Shen Q. X., Chen J., Chen L. W., Wang T. L. et al. (2000) Human mu-opioid receptor overexpressed in Sf9 insect cells functionally coupled to endogenous Gi/o proteins. Cell Res. 10: 93-102
153. Vasudevan S., Reilander H., Maul G. and Michel H. (1991) Expression and cell membrane localization of rat M3 muscarinic acetylcholine receptor produced in Sf9 insect cells using the baculovirus system. FEBS Lett. 283: 52-56
154. Hu Y., Rajan L. and Schilling W. P. (1994) Ca2+ signaling in Sf9 insect cells and the functional expression of a rat brain M5 muscarinic receptor. Am. J. Physiol. 266: C1736-1743
155. Hayashi M. K. and Haga T. (1996) Purification and functional reconstitution with GTP-binding regulatory proteins of hexahistidine-tagged muscarinic acetylcholine receptors (m2 subtype). J. Biochem. (Tokyo) 120: 1232-1238
156. Mills A., Allet B., Bernard A., Chabert C., Brandt E., Cavegn C. et al. (1993) Expression and characterization of human D4 dopamine receptors in baculovirus-infected insect cells. FEBS Lett. 320: 130-134
157. Javitch J. A., Kaback J., Li X. and Karlin A. (1994) Expression and characterization of human dopamine D2 receptor in baculovirus-infected insect cells. J. Recept. Res. 14: 99-117
158. Grunewald S., Haase W., Reilander H. and Michel H. (1996) Glycosylation, palmitoylation, and localization of the human D2S receptor in baculovirus-infected insect cells. Biochemistry 35: 15149-15161
159. Narayan P., Gray J. and Puett D. (1996) Expression of functional lutropin/choriogonadotropin receptor in the baculovirus system. Mol. Cell. Endocrinol. 117: 95-100
160. Christophe S., Robert P., Maugain S., Bellet D., Koman A. and Bidart J. M. (1993) Expression of the human follicle-stimulating hormone receptor in the baculovirus system. Biochem. Biophys. Res. Commun. 196: 402-408
161. Pettit D. A., Showalter V. M., Abood M. E. and Cabral G. A. (1994) Expression of a cannabinoid receptor in baculovirus-infected insect cells. Biochem. Pharmacol. 48: 1231-1243
162. Quehenberger O., Prossnitz E. R., Cochrane C. G. and Ye R. D. (1992) Absence of G(i) proteins in the Sf9 insect cell. Characterization of the uncoupled recombinant N-formyl peptide receptor. J. Biol. Chem. 267: 19757-19760
163. Kawamoto S., Onishi H., Hattori S., Miyagi Y., Amaya Y., Mishina M., et al. (1991) Functional expression of the alpha 1 subunit of the AMPA-selective glutamate receptor channel, using a baculovirus system. Biochem. Biophys. Res. Commun. 181: 756-763
164. Keinanen K., Kohr G., Seeburg P. H., Laukkanen M. L. and Oker-Blom C. (1994) High-level expression of functional glutamate receptor channels in insect cells. Biotechnology 12: 802-806
165. Kawamoto S., Hattori S., Oiji I., Hamajima K., Mishina M. and Okuda K. (1994) Ligand-binding properties and N-glycosylation of alpha 1 subunit of the alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionate(AMPA)-selective glutamate receptor channel expressed in a baculovirus system. Eur. J. Biochem 223: 665-673
166. Nekrasova E., Sosinskaya A., Natochin M., Lancet D. and Gat U. (1996) Overexpression, solubilization and purification of rat and human olfactory receptors. Eur. J. Biochem. 238: 28-37
167. Klaassen C. H., Bovee-Geurts P. H., Decaluwe G. L. and De-Grip W. J. (1999) Large-scale production and purification of functional recombinant bovine rhodopsin with the use of the baculovirus expression system. Biochem. J. 342: 293-300
168. Moepps B., Frodl R., Rodewald H. R., Baggiolini M. and Gierschik P. (1997) Two murine homologues of the human chemokine receptor CXCR4 mediating stromal cell-derived factor 1alpha activation of Gi2 are differentially expressed in vivo. Eur. J. Immunol. 27: 2102-2112
169. Schreurs J., Yamamoto R., Lyons J., Munemitsu S., Conroy L., Clark R., et al. (1995) Functional wild-type and carboxyl-terminal-tagged rat substance P receptors expressed in baculovirus-infected insect Sf9 cells. J. Neurochem. 64: 1622-1631
170. Nishimura K., Frederick J. and Kwatra M. M. (1998) Human substance P receptor expressed in Sf9 cells couples with multiple endogenous G proteins. J. Recept. Signal Transduct. Res. 18: 51-65
171. Beukers M. W., Klaassen C. H., De Grip W. J., Verzijl D., Timmerman H. and Leurs R. (1997) Heterologous expression of rat epitope-tagged histamine H2 receptors in insect Sf9 cells. Br. J. Pharmacol. 122: 867-874
172. Kusui T., Hellmich M. R., Wang L. H., Evans R. L., Benya R. V., Battey J. F. et al. (1995) Characterization of gastrin-releasing peptide receptor expressed in Sf9 insect cells by baculovirus. Biochemistry 34: 8061-8075
173. Ohtaki T., Ogi K., Masuda Y., Mitsuoka K., Fujiyoshi Y., Kitada C. et al. (1998) Expression, purification and reconstitution of receptor for pituitary adenylate cyclase-activating polypeptide, large-scale purification of a functionally active G protein-coupled receptor produced in Sf9 insect cells. J. Biol. Chem. 273: 15464-15473
174. Scheer A., Bjorklof K., Cotecchia S. and Lundstrom K. (1999) Expression of the alpha 1b-adrenergic receptor and G protein subunits in mammalian cell lines using the Semliki Forest virus expression system. J. Recept. Signal Transduct. Res. 19: 369-378
175. Fraser C. M., Arakawa S., McCombie W. R. and Venter J. C. (1989) Cloning, sequence analysis and permanent expression of a human alpha 2-adrenergic receptor in Chinese hamster ovary cells. Evidence for independent pathways of receptor coupling to adenylate cyclase attenuation and activation. J. Biol. Chem. 264: 11754-11761
176. Eason M. G., Kurose H., Holt B. D., Raymond J. R. and Liggett S. B. (1992) Simultaneous coupling of alpha 2-adrenergic receptors to two G-proteins with opposing effects. Subtype-selective coupling of alpha 2C10, alpha 2C4 and alpha 2C2 adrenergic receptors to Gi and Gs. J. Biol. Chem. 267: 15795-15801
177. Bresnahan M. R., Flordellis C. S., Vassilatis D. K., Makrides S. C., Zannis V. I. and Gavras H. (1990) High level of expression of functional human platelet alpha 2- adrenergic receptors in a stable mouse C127 cell line. Biochim. Biophys. Acta 1052: 439-445
178. George S. T., Berrios M., Hadcock J. R., Wang H. Y. and Malbon C. C. (1988) Receptor density and cAMP accumulation: analysis in CHO cells exhibiting stable expression of a cDNA that encodes the beta 2-adrenergic receptor. Biochem. Biophys. Res. Commun. 150: 665-672
179. Maroteaux L., Saudou F., Amlaiky N., Boschert U., Plassat J. L. and Hen R. (1992) Mouse 5HT1B serotonin receptor: cloning, functional expression and localization in motor control centers. Proc. Natl. Acad. Sci. USA 89: 3020-3024
180. McAllister G., Charlesworth A., Snodin C., Beer M. S., Noble A. J., Middlemiss D. N. et al. (1992) Molecular cloning of a serotonin receptor from human brain (5HT1E): a fifth 5HT1-like subtype. Proc. Natl. Acad. Sci. USA 89: 5517-5521
181. Saudou F., Boschert U., Amlaiky N., Plassat J. L. and Hen R. (1992) A family of Drosophila serotonin receptors with distinct intracellular signalling properties and expression patterns. EMBO J. 11: 7-17
182. Van Craenenbroeck K., Vanhoenacker P., Leysen J. E. and Haegeman G. (2001) Evaluation of the tetracycline- and ecdysone-inducible systems for expression of neurotransmitter receptors in mammalian cells. Eur. J. Neurosci. 14: 968-976
183. Lundstrom K., Mills A., Buell G., Allet E., Adami N. and Liljestrom P. (1994) High-level expression of the human neurokinin-1 receptor in mammalian cell lines using the Semliki Forest virus expression system. Eur. J. Biochem 224: 917-921
184. Turcatti G., Ceszkowski K. and Chollet A. (1993) Biochemical characterization and solubilization of human NK2 receptor expressed in Chinese hamster ovary cells. J. Recept. Res. 13: 639-652
185. Gao Z., Ni Y., Szabo G. and Linden J. (1999) Palmitoylation of the recombinant human A1 adenosine receptor: enhanced proteolysis of palmitoylation-deficient mutant receptors. Biochem. J. 342: 387-395
186. Townsend-Nicholson A. and Shine J. (1992) Molecular cloning and characterisation of a human brain A1 adenosine receptor cDNA. Brain Res. Mol. Brain Res. 16: 365-370
187. Ingi T., Kitajima Y., Minamitake Y. and Nakanishi S. (1991) Characterization of ligand-binding properties and selectivities of three rat tachykinin receptors by transfection and functional expression of their cloned cDNAs in mammalian cells. J. Pharmacol. Exp. Ther. 259: 968-975
188. Kris R. M., South V., Saltzman A., Felder S., Ricca G. A., Jaye M. et al. (1991) Cloning and expression of the human substance K receptor and analysis of its role in mitogenesis. Cell Growth Differ. 2: 15-22
189. Nakajima Y., Tsuchida K., Negishi M., Ito S. and Nakanishi S. (1992) Direct linkage of three tachykinin receptors to stimulation of both phosphatidylinositol hydrolysis and cyclic AMP cascades in transfected Chinese hamster ovary cells. J. Biol. Chem. 267: 2437-2442
190. Wang J. B., Imai Y., Eppler C. M., Gregor P., Spivak C. E. and Uhl G. R. (1993) mu opiate receptor: cDNA cloning and expression. Proc. Natl. Acad. Sci. USA 90: 10230-10234
191. Raynor K., Kong H., Mestek A., Bye L. S., Tian M., Liu J. et al. (1995) Characterization of the cloned human mu opioid receptor. J. Pharmacol. Exp. Ther. 272: 423-428
192. Peralta E. G., Ashkenazi A., Winslow J. W., Ramachandran J. and Capon D. J. (1988) Differential regulation of PI hydrolysis and adenylyl cyclase by muscarinic receptor subtypes. Nature 334: 434-437
193. Migeon J. C. and Nathanson N. M. (1994) Differential regulation of cAMP-mediated gene transcription by m1 and m4 muscarinic acetylcholine receptors. Preferential coupling of m4 receptors to Gi alpha-2. J. Biol. Chem. 269: 9767-9773
194. Schweitzer C., Kratzeisen C., Adam G., Lundstrom K., Malherbe P., Ohresser S. et al. (2000) Characterization of [(3)H]-LY354740 binding to rat mGlu2 and mGlu3 receptors expressed in CHO cells using Semliki Forest virus vectors. Neuropharmacology 39: 1700-1706
195. Malherbe P., Kratzeisen C., Lundstrom K., Richards J. G., Faull R. L. and Mutel V. (1999) Cloning and functional expression of alternative spliced variants of the human metabotropic glutamate receptor α. Brain Res. Mol. Brain Res. 67: 201-210
196. Walker P., Munoz M., Combe M. C., Grouzmann E., Herzog H., Selbie L. et al. (1993) High level expression of human neuropeptide Y receptors in mammalian cells infected with a recombinant vaccinia virus. Mol. Cell. Endocrinol. 91: 107-112
197. Falck-Pedersen E., Heinflink M., Alvira M., Nussenzveig D. R. and Gershengorn M. C. (1994) Expression of thyrotropin-releasing hormone receptors by adenovirus-mediated gene transfer reveals that thyrotropin-releasing hormone desensitization is cell specific. Mol. Pharmacol. 45: 684-689
198. Jelinek L. J., Lok S., Rosenberg G. B., Smith R. A., Grant F. J., Biggs S. et al. (1993) Expression cloning and signaling properties of the rat glucagon receptor. Science 259: 1614-1616
199. Needham M., Egerton M., Millest A., Evans S., Popplewell M., Cerillo G. et al. (1995) Further development of the locus control region/murine erythroleukemia expression system: high level expression and characterization of recombinant human calcitonin receptor. Protein Expr. Purif. 6: 124-131
200. Pawate S., Schey K. L., Meier G. P., Ullian M. E., Mais D. E. and Halushka P. V. (1998) Expression, characterization and purification of C-terminally hexahistidine-tagged thromboxane A2 receptors. J. Biol. Chem. 273: 22753-22760
201. Elshourbagy N. A., Lee J. A., Korman D. R., Nuthalaganti P., Sylvester D. R., Dilella A. G. et al. (1992) Molecular cloning and characterization of the major endothelin receptor subtype in porcine cerebellum. Mol. Pharmacol. 41: 465-473
202. Hoffmann M., Verzijl D., Lundstrom K., Simmen U., Alewijnse A. E., Timmerman H. et al. (2001) Recombinant Semliki Forest virus for over-expression and pharmacological characterisation of the histamine H(2) receptor in mammalian cells. Eur. J. Pharmacol. 427: 105-114
203. Hatt H., Lang K. and Gisselmann G. (2001) Functional expression and characterization of odorant receptors using the Semliki Forest virus system. Biol. Chem. 382: 1207-1214
204. Oprian D. D., Molday R. S., Kaufman R. J. and Khorana H. G. (1987) Expression of a synthetic bovine rhodopsin gene in monkey kidney cells. Proc. Natl. Acad. Sci. USA 84: 8874-8878
205. Baneres J. L., Martin A. et al. (2003) "Structure-based analysis of GPCR function: conformational adaptation of both agonist and receptor upon leukotriene B4 binding to recombinant BLT1." J Mol Biol 329(4): 801-814
206. Baneres J. L. and Parello J. (2003) "Structure-based analysis of GPCR function: evidence for a novel pentameric assembly between the dimeric leukotriene B4 receptor BLT1 and the G-protein." J Mol Biol 329(4): 815-829.
207. Kwatra M. M., Schreurs J. et al. (1995) "Immunoaffinity purification of epitope-tagged human beta 2-adrenergic receptor to homogeneity." Protein Expr Purif 6(6): 717-721
208. Price L. A., Strnad J. et al. (1996). "Pharmacological characterization of the rat A2a adenosine receptor funcitonally coupled to the yeast pheromone response pathway." Mol Pharmacol 50(4): 829-837