Site-Specific Labeling of Genetically Encoded Azido Groups for Multicolor, Single-Molecule Fluorescence Imaging of GPCRs

Methods in Cell Biology

Volumn 117, Issue , 2013, Pages 267-303

  Tian, He ^a   Sakmar, Thomas P ^a   Huber, Thomas ^a  

^a ROCKEFELLER UNIVERSITY   (United States)

Author keywords

Alkyne azide cycloaddition; Bioorthogonal ligation; Fluorophore photostability; G protein coupled receptors; Mammalian cells; Single molecule fluorescence; Site specific incorporation; Structural dynamics; Surface chemistry; Transfer RNA synthetase; Transmembrane proteins; Unnatural amino acid

Indexed keywords

ALEXA 488 HYDRAZIDE; AZIDE; FLUORESCENT DYE; HYDRAZINE DERIVATIVE; OPSIN; RETINAL; RHODOPSIN;

ALKYNE-AZIDE CYCLOADDITION; ARTICLE; BIOORTHOGONAL LIGATION; CHEMISTRY; CYCLOADDITION; FLUOROPHORE PHOTOSTABILITY; G PROTEIN-COUPLED RECEPTORS; GENE EXPRESSION; GENETIC TRANSFECTION; GENETICS; HEK293 CELL LINE; HUMAN; IMMUNOPRECIPITATION; KINETICS; MAMMALIAN CELLS; METABOLISM; METHODOLOGY; MOLECULAR IMAGING; PROTEIN MULTIMERIZATION; SIGNAL TRANSDUCTION; SINGLE-MOLECULE FLUORESCENCE; SITE-SPECIFIC INCORPORATION; SPECTROFLUOROMETRY; STAINING; STOP CODON; STRUCTURAL DYNAMICS; SURFACE CHEMISTRY; TRANSFER-RNA SYNTHETASE; TRANSMEMBRANE PROTEINS; UNNATURAL AMINO ACID;

ALKYNE-AZIDE CYCLOADDITION; BIOORTHOGONAL LIGATION; FLUOROPHORE PHOTOSTABILITY; G PROTEIN-COUPLED RECEPTORS; MAMMALIAN CELLS; SINGLE-MOLECULE FLUORESCENCE; SITE-SPECIFIC INCORPORATION; STRUCTURAL DYNAMICS; SURFACE CHEMISTRY; TRANSFER-RNA SYNTHETASE; TRANSMEMBRANE PROTEINS; UNNATURAL AMINO ACID;

AZIDES; CODON, NONSENSE; CYCLOADDITION REACTION; FLUORESCENT DYES; GENE EXPRESSION; HEK293 CELLS; HUMANS; HYDRAZINES; IMMUNOPRECIPITATION; KINETICS; MOLECULAR IMAGING; OPSINS; PROTEIN MULTIMERIZATION; RETINALDEHYDE; RHODOPSIN; SIGNAL TRANSDUCTION; SPECTROMETRY, FLUORESCENCE; STAINING AND LABELING; TRANSFECTION;

EID: 84885936031     PISSN: 0091679X     EISSN: None     Source Type: Book Series    
DOI: 10.1016/B978-0-12-408143-7.00015-3     Document Type: Chapter

References (20)

1. Axelrod D., Burghardt T.P., Thompson N.L. Total internal-reflection fluorescence. Annual Review of Biophysics and Bioengineering 1984, 13:247-268.
2. Blanchard S.C. Single-molecule observations of ribosome function. Current Opinion in Structural Biology 2009, 19:103-109.
3. Cordes T., Maiser A., Steinhauer C., Schermelleh L., Tinnefeld P. Mechanisms and advancement of antifading agents for fluorescence microscopy and single-molecule spectroscopy. Physical Chemistry Chemical Physics 2011, 13:6699-6709.
4. Franke R.R., Sakmar T.P., Oprian D.D., Khorana H.G. A single amino-acid substitution in rhodopsin (lysine-248→leucine) prevents activation of transducin. Journal of Biological Chemistry 1988, 263:2119-2122.
5. Huber T., Sakmar T.P. Escaping the flatlands: New approaches for studying the dynamic assembly and activation of GPCR signaling complexes. Trends in Pharmacological Sciences 2011, 32:410-419.
6. Joo C., Balci H., Ishitsuka Y., Buranachai C., Ha T. Advances in single-molecule fluorescence methods for molecular biology. Annual Review of Biochemistry 2008, 77:51-76.
7. Kapanidis A.N., Laurence T.A., Lee N.K., Margeat E., Kong X.X., Weiss S. Alternating-laser excitation of single molecules. Accounts of Chemical Research 2005, 38:523-533.
8. Knepp A.M., Grunbeck A., Banerjee S., Sakmar T.P., Huber T. Direct measurement of thermal stability of expressed CCR5 and stabilization by small molecule ligands. Biochemistry 2011, 50:502-511.
9. Naganathan S., Ye S.X., Sakmar T.P., Huber T. Site-specific epitope tagging of G protein-coupled receptors by bioorthogonal modification of a genetically encoded unnatural amino acid. Biochemistry 2013, 52:1028-1036.
10. Oprian D.D., Molday R.S., Kaufman R.J., Khorana H.G. Expression of a synthetic bovine rhodopsin gene in monkey kidney-cells. Proceedings of the National Academy of Sciences of the United States of America 1987, 84:8874-8878.
11. Rolfe D.J., McLachlan C.I., Hirsch M., Needham S.R., Tynan C.J., Webb S.E.D., et al. Automated multidimensional single molecule fluorescence microscopy feature detection and tracking. European Biophysics Journal 2011, 40:1167-1186.
12. Roy R., Hohng S., Ha T. A practical guide to single-molecule FRET. Nature Methods 2008, 5:507-516.
13. Sage D., Neumann F.R., Hediger F., Gasser S.M., Unser M. Automatic tracking of individual fluorescence particles: Application to the study of chromosome dynamics. IEEE Transactions on Image Processing 2005, 14:1372-1383.
14. Sakmar T.P., Franke R.R., Khorana H.G. Glutamic acid-113 serves as the retinylidene Schiff-base counterion in bovine rhodopsin. Proceedings of the National Academy of Sciences of the United States of America 1989, 86:8309-8313.
15. Serge A., Bertaux N., Rigneault H., Marguet D. Dynamic multiple-target tracing to probe spatiotemporal cartography of cell membranes. Nature Methods 2008, 5:687-694.
16. Swoboda M., Henig J., Cheng H.M., Brugger D., Haltrich D., Plumere N., et al. Enzymatic oxygen scavenging for photostability without pH drop in single-molecule experiments. ACS Nano 2012, 6:6364-6369.
17. Vogelsang J., Kasper R., Steinhauer C., Person B., Heilemann M., Sauer M., et al. A reducing and oxidizing system minimizes photobleaching and blinking of fluorescent dyes. Angewandte Chemie International Edition 2008, 47:5465-5469.
18. Weiss S. Fluorescence spectroscopy of single biomolecules. Science 1999, 283:1676-1683.
19. Ye S.X., Huber T., Vogel R., Sakmar T.P. FTIR analysis of GPCR activation using azido probes. Nature Chemical Biology 2009, 5:397-399.
20. Ye S.X., Kohrer C., Huber T., Kazmi M., Sachdev P., Yan E.C.Y., et al. Site-specific incorporation of keto amino acids into functional G protein-coupled receptors using unnatural amino acid mutagenesis. Journal of Biological Chemistry 2008, 283:1525-1533.