Adding value to fusion proteins through covalent labelling

Current Opinion in Biotechnology

Volumn 16, Issue 4, 2005, Pages 453-458

  Gronemeyer, Thomas ^a   Godin, Guillaume ^a   Johnsson, Kai ^a  

^a EPFL   (Switzerland)

Author keywords

[No Author keywords available]

Indexed keywords

COVALENT LABELING; LIVING CELLS; PROTEIN SCIENCE; RECOMBINANT PROTEINS;

BIOTECHNOLOGY; CELLS; GENETIC ENGINEERING; POLYPEPTIDES; PURIFICATION;

PROTEINS;

6 O ALKYLGUANINE DNA ALKYLTRANSFERASE; ADENOSINE A2 RECEPTOR; CYSTEINE; FLUORESCEIN; G PROTEIN COUPLED RECEPTOR; HYBRID PROTEIN; RESORUFIN DERIVATIVE;

CARBOXY TERMINAL SEQUENCE; COVALENT BOND; DEHALOGENATION; ENZYME SUBSTRATE; FLUORESCENCE RESONANCE ENERGY TRANSFER; HAPLOIDY; PRIORITY JOURNAL; PROTEIN EXPRESSION; PROTEIN MOTIF; PROTEIN PROCESSING; PROTEIN PROTEIN INTERACTION; PROTEIN STABILITY; PROTEIN STRUCTURE; PROTEIN SYNTHESIS; REVIEW; SACCHAROMYCES CEREVISIAE; STRUCTURE ANALYSIS;

RECOMBINANT FUSION PROTEINS; STAINING AND LABELING;

EID: 23444448243     PISSN: 09581669     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.copbio.2005.06.001     Document Type: Review

References (29)

1. L. Wang, and P.G. Schultz Expanding the genetic code Angew Chem Int Ed Engl 44 2004 34 66
2. N. Johnsson, and K. Johnsson A fusion of disciplines: chemical approaches to exploit fusion proteins for functional genomics ChemBioChem 4 2003 803 810
3. B.A. Griffin, S.R. Adams, and R.Y. Tsien Specific covalent labeling of recombinant protein molecules inside live cells Science 281 1998 269 272
4. S.R. Adams, R.E. Campbell, L.A. Gross, B.R. Martin, G.K. Walkup, Y. Yao, J. Llopis, and R.Y. Tsien New biarsenical ligands and tetracysteine motifs for protein labeling in vitro and in vivo: synthesis and biological applications J Am Chem Soc 124 2002 6063 6076
5. G. Gaietta, T.J. Deerinck, S.R. Adams, J. Bouwer, O. Tour, D.W. Laird, G.E. Sosinsky, R.Y. Tsien, and M.H. Ellisman Multicolor and electron microscopic imaging of connexin trafficking Science 296 2002 503 507 Sequential multicolor labelling of fusion proteins of connexin and the tetracysteine tag is used here to study the dynamic behavior of connexin. ReAsH-labelled connexin fusion proteins were also used for the localized photoconversion of diaminobenzidine into an electron-dense precipitate. Subsequent high-resolution electron microscopy imaging demonstrated the multifunctional aspect of the tag.
6. M. Andresen, R. Schmitz-Salue, and S. Jakobs Short tetracysteine tags to β-tubulin demonstrate the significance of small labels for live cell imaging Mol Biol Cell 15 2004 5616 5622
7. C. Hoffmann, G. Gaietta, M. Bünemann, S.R. Adams, S. Oberdorff-Maass, B. Behr, J.P. Vilardaga, R.Y. Tsien, M.H. Ellisman, and M.J. Lohse A FlAsH-based FRET approach to measure G protein-coupled receptor activation in living cells Nat Methods 2 2005 171 176 The authors demonstrate that FRET from CFP to FlAsH in GPCR fusion proteins can be used to monitor GPCR activation in living cells without disturbing receptor function. This is in contrast to experiments based on FRET between CFP and YFP and demonstrates that the small size of the tetracysteine tag can be decisively advantageous.
8. R.Y. Tsien Building and breeding molecules to spy on cells and tumors FEBS Lett 579 2005 927 932
9. E.G. Guignet, R. Hovius, and H. Vogel Reversible site-selective labeling of membrane proteins in live cells Nat Biotechnol 22 2004 440 444 A method is presented for the labelling of oligohistidine fusion proteins with synthetic probes using metal ion chelating NTA derivatives. The labelling is very fast and reversible and applications are shown for functional studies of GPCRs. A possible drawback of the approach is modest selectivity of the labelling and stability of the construct.
10. A. Keppler, S. Gendreizig, T. Gronemeyer, H. Pick, H. Vogel, and K. Johnsson A general method for the covalent labeling of fusion proteins with small molecules in vivo Nat Biotechnol 21 2003 86 89
11. 6-alkylguanine by alkyltransferases Mutat Res 462 2000 83 100
12. A. Keppler, H. Pick, C. Arrivoli, H. Vogel, and K. Johnsson Labeling of fusion proteins with synthetic fluorophores in live cells Proc Natl Acad Sci USA 101 2004 9955 9959 The sequential labelling of AGT fusion proteins with different fluorophores in mammalian cells is described. AGT fusion proteins with different localizations in the cell can be specifically labelled with different fluorophores and the fluorescence-labelling used for applications such as multicolor analysis of dynamic processes and FRET measurements, demonstrating the general applicability of the approach.
13. S. Gendreizig, M. Kindermann, and K. Johnsson Induced protein dimerization in vivo through covalent labeling J Am Chem Soc 125 2003 14970 14971
14. M. Kindermann, N. George, N. Johnsson, and K. Johnsson Covalent and selective immobilization of fusion proteins J Am Chem Soc 125 2003 7810 7811
15. M. Kindermann, I. Sielaff, and K. Johnsson Synthesis and characterization of bifunctional probes for the specific labeling of fusion proteins Bioorg Med Chem Lett 14 2004 2725 2728
16. 6-alkylguanine-DNA alkyltransferase for efficient labeling of fusion proteins with small molecules in vivo Chem Biol 10 2003 313 317
17. 6-alkylguanine- DNA alkyltransferase for specific protein labeling in living cells. ChemBioChem 2005, 7: in press.
18. D was in excellent agreement with that measured for cyclophilin in solution, demonstrating that the protein is immobilized in its native state.
19. L.W. Miller, J. Sable, P. Goelet, M.P. Sheetz, and V.W. Cornish Methotrexate conjugates: a molecular in vivo protein tag Angew Chem Int Ed Engl 43 2004 1672 1675
20. K.M. Marks, P.D. Braun, and G.P. Nolan A general approach for chemical labeling and rapid, spatially controlled protein inactivation Proc Natl Acad Sci USA 101 2004 9982 9987
21. J.J. La Clair, T.L. Foley, T.R. Schegg, C.M. Regan, and M.D. Burkart Manipulation of carrier proteins in antibiotic biosynthesis Chem Biol 11 2004 195 201
22. •].
23. •] and the labelling of cell-surface proteins with fluorophores, quantum dots and affinity probes. The labelling is fast and highly specific; a potential pitfall of the approach is modification of the CP fusion proteins by the PPTase of the expression host.
24. N. Johnsson, N. George, and K. Johnsson Protein chemistry on the surface of living cells ChemBioChem 6 2005 47 52
25. J. Yin, F. Liu, M. Schinke, C. Daly, and C.T. Walsh Phagemid encoded small molecules for high throughput screening of chemical libraries J Am Chem Soc 126 2004 13570 13571
26. J.E. Cronan Jr. Biotination of proteins in vivo. A post-translational modification to label, purify, and study proteins J Biol Chem 265 1990 10327 10333
27. D. Beckett, E. Kovaleva, and P.J. Schatz A minimal peptide substrate in biotin holoenzyme synthetase-catalyzed biotinylation Protein Sci 8 1999 921 929
28. I. Chen, M. Howarth, W. Lin, and A.Y. Ting Site-specific labeling of cell surface proteins with biophysical probes using biotin ligase Nat Methods 2 2005 99 104 It is demonstrated in this paper that BirA specifically ligates a ketone isostere of biotin to cell-surface fusion proteins displaying biotin acceptor peptides; the cell-surface fusion protein can subsequently be labelled with hydrazides carrying biophysical probes. An advantage of the approach is the small size of the tag; however, a limitation is the slow reaction between the ketone and the hydrazide.
29. D.A. Nauman, and C.R. Bertozzi Kinetic parameters for small-molecule drug delivery by covalent cell surface targeting Biochim Biophys Acta 1568 2001 147 154