Long-Lifetime Metal—Ligand Complexes

  • Joseph R. Lakowicz

Abstract

Throughout the previous chapters, we described fluorophores with decay times ranging from 1 to 20 ns. While this timescale is useful for many biophysical measurements, there are numerous instances where longer decay times are desirable. For instance, one may wish to measure rotational motions of large proteins or membrane-bound proteins. In such cases the overall rotational correlation times can easily be longer than 200 ns, and they can exceed 1 μs for larger macromolecular assemblies. Rotational motions on this timescale are not measurable using fluorophores which display nanosecond lifetimes. Processes on the microsecond or even the millisecond timescale have occasionally been measured using phosphorescence.1–4 However, relatively few probes display useful phosphorescence in room-temperature aqueous solutions. Also, it is usually necessary to perform phosphorescence measurements in the complete absence of oxygen. Hence, there is a clear need for probes which display microsecond lifetimes. In this chapter we describe a family of metal—ligand probes which display decay times ranging from 100 ns to 10μs. The long lifetimes of the metal—ligand probes allow the use of gated detection, which can be employed to suppress interfering autofluorescence from biological samples and can thus provide increased sensitivity.5 Finally, the metal—ligand probes display high chemical and photochemical stability. Because of these favorable properties, we expect metal—ligand probes to have numerous uses in biophysical chemistry, clinical chemistry, and DNA diagnostics.

Keywords

Human Serum Albumin Decay Time Rotational Correlation Time Intensity Decay Fluorescence Polarization Immunoassay 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Vanderkooi, J. M., 1992, Tryptophan phosphorescence from proteins at room temperature, in Topics in Fluorescence Spectroscopy. Volume 3, Biochemical Applications, J. R. Lakowicz (ed.), Plenum Press, New York, pp. 113–136.Google Scholar
  2. 2.
    Hurtubise, R. J., 1990, Phosphorimetry: Theory, Instrumentation, and Applications, VCH Publishers, New York.Google Scholar
  3. 3.
    Barthold, M., Barrantes, F. J., and Jovin, T. M., 1981, Rotational molecular dynamics of the membrane-bound acetylcholine receptor revealed by phosphorescence spectroscopy, Eur. J. Biochem. 120: 389–397.CrossRefGoogle Scholar
  4. 4.
    Che, A., and Cherry, R. J., 1995, Loss of rotational mobility of band 3 proteins in human erythrocyte membranes induced by antibodies to glycophorin A, Biophys. J. 68: 1881–1887.CrossRefGoogle Scholar
  5. 5.
    Haugen, G. R., and Lytle, F. E., 1981, Quantitation of fluorophores in solution by pulsed laser excitation and time-filtered detection, Anal. Chem. 53: 1554–1559.CrossRefGoogle Scholar
  6. 6.
    Juris, A., Balzani, V., Barigelletti, F., Campagna, S., Belser, P., and Von Zelewsky, A., 1988, Ru(II) polypyridine complexes: Photo-physics, photochemistry, electrochemistry and chemiluminescence, Coord. Chem. Rev. 84: 85–277.CrossRefGoogle Scholar
  7. 7.
    Demas, J. N., and DeGraff, B. A., 1994, Design and applications of highly luminescent transition metal complexes, in Topics in Fluorescence Spectroscopy, Volume 4, Probe Design and Chemical Sensing, J. R. Lakowicz (ed.), Plenum Press, New York, pp. 71–107.Google Scholar
  8. 8.
    F. N. Castellano, personal communication.Google Scholar
  9. 9.
    Damrauer, N. H., Cerullo, G., Yeh, A., Boussie, T. R., Shamk, C. V., and McCusker, J. K., 1997, Femtosecond dynamics of excited state evolution in [Ru(bpy)3]2+, Science 275: 54–57.CrossRefGoogle Scholar
  10. 10.
    Demas, J. N., and DeGraff, B. A., 1991, Design and applications of highly luminescent transition metal complexes. Anal. Chem. 63: 829A - 837A.Google Scholar
  11. 11.
    Kalayanasundaram, K., 1992, Photochemistry of Polypyridine and Porphyrin Complexes, Academic Press, New York.Google Scholar
  12. 12.
    Demas, J. N., and DeGraff, B. A., 1997, Applications of luminescent transition metal complexes to sensor technology and molecular probes, J. Chem. Educ. 74: 690–695.CrossRefGoogle Scholar
  13. 13.
    Terpetschnig, E., Szmacinski, H., Malak., H., and Lakowicz, J. R., 1995, Metal-ligand complexes as a new class of long-lived fluorophores for protein hydrodynamics, Biophys. J. 68: 342–350.Google Scholar
  14. 14.
    Szmacinski, H., Terpetschnig, E., and Lakowicz, J.R., 1996, Synthesis and evaluation of Ru-complexes as anisotropy probes for protein hydrodynamics and immunoassays of high-molecular-weight antigens, Biophys. Chem. 62: 109–120.CrossRefGoogle Scholar
  15. 15.
    Castellano, F. N., Dattelbaum, J. D., and Lakowicz, J. R., 1998, Long-lifetime Ru(II) complexes as labeling reagents for sulfhydryl groups, Anal. Biochem. 255: 165–170.CrossRefGoogle Scholar
  16. 16.
    Terpetschnig, E., Dattelbaum, J. D., Szmacinski, H., and Lakowicz, J. R., 1997, Synthesis and spectral characterization of a thiol-reactive long-lifetime Ru(II) complex, Anal. Biochem. 251: 241–245.CrossRefGoogle Scholar
  17. 17.
    Guo X., Li, L., Castellano, E. N., Szmacinski, H., and Lakowicz, J. R., 1997, A long-lived, highly luminescent rhenium (I) metal-ligand complex as a bimolecular probe, Anal. Biochem. 254: 179–186.CrossRefGoogle Scholar
  18. 18.
    Terpetschnig, E., Szmacinski, H., and Lakowicz, J. R., 1996, Fluorescence polarization immunoassay of a high molecular weight antigen using a long wavelength absorbing and laser diode-excitable metal-ligand complex, Anal. Biochem. 240: 54–59.CrossRefGoogle Scholar
  19. 19.
    Lakowicz, J. R., Terpetschnig, E., Murtaza, Z., and Szmacinski, H., 1997, Development of long-lifetime metal-ligand probes for biophysics and cellular imaging, J. Fluoresc. 7 (1): 17–25.CrossRefGoogle Scholar
  20. 20.
    Demas, J. N., and DeGraff, B. A., 1992, Applications of highly luminescent transition metal complexes in polymer systems, Makromol. Chem., MacromoL Symp. 59: 35–51.CrossRefGoogle Scholar
  21. 21.
    Caspar, J., and Meyer, T. J., 1983, Application of the energy gap law to nonradiative, excited-state decay, J. Phys. Chem. 87: 952–957.CrossRefGoogle Scholar
  22. 22.
    Kober, E. M., Marshall, J. L., Dressick, W. J., Sullivan, B. P., Caspar, J. V., and Meyer, T. J., 1985, Synthetic control of excited states. Nonchromophoric ligand variations in polypyridyl complexes of osmium(II), lnorg. Chem. 24: 2755–2763.CrossRefGoogle Scholar
  23. 23.
    Kober, E. M., Sullivan, B. P., Dressick, W. J., Caspar, J. V, and Meyer, T. J., 1980, Highly luminescent polypyridyl complexes of osmium(II), J. Am. Chem. Soc. 102: 1383–1385.CrossRefGoogle Scholar
  24. 24.
    Caspar, J. V, and Meyer, T. J., 1983, Photochemistry of Ru(bpy)3+solvent effects, J. Am. Chem. Soc. 105: 5583–5590.CrossRefGoogle Scholar
  25. 25.
    Caspar, J. V., Kober, E. M., Sullivan, B. P., and Meyer, T. J., 1982, Application of the energy gap law to the decay of charge-transfer excited states, J. Am. Chem. Soc. 104: 630–632.CrossRefGoogle Scholar
  26. 26.
    Bixon, M., and Jortner, J., 1968, Intramolecular radiationless transitions, J. Chem. Phys. 48: 715–726.CrossRefGoogle Scholar
  27. 27.
    Freed, K. F., and Jortner, J., 1970, Multiphonon processes in the nonradiative decay of large molecules, J. Chem. Phys. 52: 6272–6291.CrossRefGoogle Scholar
  28. 28.
    Barkley, M. D., and Zimm, B. H., 1979, Theory of twisting and bending of chain macromolecules: Analysis of the fluorescence depolarization of DNA, J. Chem. Phys. 70: 2991–3007.CrossRefGoogle Scholar
  29. 29.
    Schurr, J. M., Fujimoto, B. S., Wu, P., and Song, L., 1992, Fluorescence studies of nucleic acids: Dynamics, rigidities and structures, in Topics in Fluorescence Spectroscopy, Volume 3, Biochemical Applications, J. R. Lakowicz (ed.), Plenum Press, New York, pp. 137–229.Google Scholar
  30. 30.
    Genest, D., Wahl, P., Erard, M., Champagne, M., and Daune, M., 1982, Fluorescence anisotropy decay of ethidium bromide bound to nucleosomal core particles, Biochimie 64: 419–427.CrossRefGoogle Scholar
  31. 31.
    Millar, D. P., Robbins, R. J., and Zewail, A. H., 1980, Direct observation of the torsional dynamics of DNA and RNA by picosecond spectroscopy, Proc. Natl. Acad. Sci. U.S.A. 77: 5593–5597.CrossRefGoogle Scholar
  32. 32.
    Millar, D. P., Robbins, R. J., and Zewail, A. H., 1982, Torsional and bending of nucleic acids studied by subnanosecond time-resolved fluorescence depolarization of intercalated dyes, J. Chem. Phys. 76: 2080–2094.CrossRefGoogle Scholar
  33. 33.
    Friedman, A. E., Chambron, J.-C., Sauvage, J.-P., Turro, N. J., and Barton, J. K., 1990, Molecular light switch for DNA Ru(bpy)2(dppz)2+, J. Am. Chem. Soc. 112: 4960–4962.CrossRefGoogle Scholar
  34. 34.
    Jenkins, Y., Friedman, A. E., Turro, N. J., and Barton, J. K., 1992, Characterization of dipyridophenazine complexes of ruthenium(II): The light switch effect as a function of nucleic acid sequence and conformation, Biochemistry 31: 10809–10816.CrossRefGoogle Scholar
  35. 35.
    Holmlin, R. E., Stemp, E. D. A., and Barton, J. K., 1998, Ru(phen)2dppz2+ luminescence: Dependence on DNA sequences and groove-binding agents, Inorg. Chem. 37: 29–34.CrossRefGoogle Scholar
  36. 36.
    Chang, Q., Murtaza, Z., Lakowicz, J. R., and Rao, G., 1997, A fluorescence lifetime-based solid sensor for water, Anal. Chim. Acta 350: 97–104.CrossRefGoogle Scholar
  37. 37.
    Guo, X-Q., Castellano, E. N., Li, L., and Lakowicz, J. R., 1998, A long-lifetime Ru(II) metal-ligand complex as a membrane probe, Biophys. Chem. 71: 51–62.CrossRefGoogle Scholar
  38. 38.
    Lakowicz, J. R., Malak, H., Gryczynski, I., Castellano, E N., and Meyer, G. J., 1995, DNA dynamics observed with long lifetime metal-ligand complexes, Biospectroscopy 1: 163–168.CrossRefGoogle Scholar
  39. 39.
    Malak, H., Gryczynski, I., Lakowicz, J. R., Meyer, G. J., and Castellano, F. N., 1997, Long-lifetime metal-ligand complexes as luminescent probes for DNA, J. Fluoresc. 7 (2): 107–112.CrossRefGoogle Scholar
  40. 40.
    Gerstein, M., Lesk, A. M., and Chothia, C., 1994, Structural mechanisms for domain movements in proteins, Biochemistry 33: 6739–6749.CrossRefGoogle Scholar
  41. 41.
    Anderson, C. M., Zucker, E H., and Steitz, T. A., 1979, Space-filling models of kinase clefts and conformation changes, Science 204: 375–380.CrossRefGoogle Scholar
  42. 42.
    Grossman, S. H. 1990, Resonance energy transfer between the active sites of creatine kinase from rabbit brain, Biochem. Biophys. Acta 1040: 276–280.CrossRefGoogle Scholar
  43. 43.
    Grossman, S. H. 1989, Resonance energy transfer between the active sites of rabbit muscle creatine kinase: Analysis by steady-state and time-resolved fluorescence, Biochemistry 28: 5902–5908.CrossRefGoogle Scholar
  44. 44.
    Haran, G., Haas, E., Szpikowska, B. K., and Mas, M. T., 1992, Domain motions in phosphoglycerate kinase: Determination of interdomain distance-distributions by site-specific labeling and time-resolved fluorescence energy transfer, Proc. Natl. Acad. Sci. U.S.A. 89: 11764–11768.CrossRefGoogle Scholar
  45. 45.
    Holowka, D., Wensel, T., and Baird, B., 1990, A nanosecond fluorescence depolarization study on the segmental flexibility of receptor-bound immunoglobulin E, Biochemistry 29: 4607–4612.CrossRefGoogle Scholar
  46. 46.
    Zheng, Y., Shopes, B., Holowka, D., and Biard, B., 1991, Conformations of IgE bound to its receptor Fc epsilon RI and in solution, Biochemistry 30: 9125–9132.CrossRefGoogle Scholar
  47. 47.
    Davenport, L., and Targowski, P., 1996, Submicrosecond phospholipid dynamics using a long-lived fluorescence emission anisotropy probe, Biophys. J. 71: 1837–1852.CrossRefGoogle Scholar
  48. 48.
    Doring, K., Beck, W., Konermann, L., and Jahnig, E, 1997, The use of a long-lifetime component of tryptophan to detect slow orientational fluctuations of proteins, Biophys. J. 72: 326–334.CrossRefGoogle Scholar
  49. 49.
    Li, L., Szmacinski, H., and Lakowicz, J. R., 1997, Long-lifetime lipid probe containing a luminescent metal-ligand complex, Biospectroscopy 3 (2): 155–159.CrossRefGoogle Scholar
  50. 50.
    Li, L., Szmacinski, H., and Lakowicz, J.R., 1997, Synthesis and luminescence spectral characterization of long-lifetime lipid metal-ligand probes, Anal. Biochem. 244: 80–85.CrossRefGoogle Scholar
  51. 51.
    Terpetschnig, E., Szmacinski, H., and Lakowicz, J. R., 1995, Fluorescence polarization immunoassay of a high molecular weightGoogle Scholar
  52. antigen based on a long-lifetime Ru—ligand complex, Anal. Biochem. 227:140–147.Google Scholar
  53. 52.
    Youn, H. J., Terpetschnig, E., Szmacinski, H., and Lakowicz, J.R., 1995, Fluorescence energy transfer immunoassay based on a long-lifetime luminescence metal—ligand complex, Anal. Biochem. 232: 24–30.CrossRefGoogle Scholar
  54. 53.
    Guo, X-Q., Castellano, E. N., Li, L., and Lakowicz, J. R., 1998, Use of a long-lifetime Re(I) complex in fluorescence polarization immunoassays of high-molecular weight analytes, Anal. Chem. 70: 632–637.CrossRefGoogle Scholar
  55. 54.
    Sipior, J., Carter, G. M., Lakowicz, J. R., and Rao, G., 1997, A blue light-emitting diode demonstrated as an ultraviolet excitation source for nanosecond phase-modulation fluorescence lifetime measurements, Rev. Sci. Instrum. 68: 2666–2670.CrossRefGoogle Scholar
  56. 55.
    Lakowicz, J. R., Murtaza, Z., Jones, W. E., Kim, K., and Szmacinski, H., 1996, Polarized emission from a rhenium metal—ligand complex, J. Fluoresc. 6 (4): 245–249.CrossRefGoogle Scholar
  57. 56.
    Brewer, R. G., Jensen, G. E., and Brewer, K. J., 1994, Long-lived osmium (II) chromophores containing 2,3,5,6-tetrakis(2pyridyl)pyrazine, Inorg. Chem. 33: 124–129.CrossRefGoogle Scholar
  58. 57.
    Murtaza, Z., and Lakowicz, J. R., 1999, Long-lifetime and long-wavelength osmium(II) metal compounds containing polypyridine ligands. Excellent red fluorescent dyes for biophysics and sensors, SPIE 3602, in press.Google Scholar
  59. 58.
    Murtaza, Z., Chang, Q., Rao, G., Lin, H., and Lakowicz, J. R., 1997, Long-lifetime metal—ligand pH probe, Anal. Biochem. 247: 216–222.CrossRefGoogle Scholar
  60. 59.
    Szmacinski, H., and Lakowicz, J. R., 1994, Lifetime-based sensing, in Topics in Fluorescence Spectroscopy, Volume 4, Probe Design and Chemical Sensing, J. R. Lakowicz (ed.), Plenum Press, New York, pp. 295–334.Google Scholar
  61. 60.
    Medicare, Medicaid and CLIA programs; Regulations implementing the clinical laboratory improvement amendments of 1988 (CLIA ‘88), 1992, Federal Register 57: 7002–7018.Google Scholar
  62. 61.
    Shen, Y., and Sullivan, B. P., 1995, A versatile preparative route to 5-substituted-1,10-phenanthroline ligands via 1,10-phenanthroline 5,6-epoxide, lnorg. Chem. 34: 6235–6236.CrossRefGoogle Scholar
  63. 62.
    Shen, Y., and Sullivan, B. P., 1997, Luminescence sensors for cations based on designed transition metal complexes, J. Chem. Educ. 74: 685–689.CrossRefGoogle Scholar
  64. 63.
    Fujita, E., Milder, S. J., and Brunschwig, B. S., 1992, Photophysical properties of covalently attached Ru(bpy)3+ and Mcyclam4+ (M = Ni, H2) complexes, Inorg. Chem. 31: 2079–2085.CrossRefGoogle Scholar
  65. 64.
    Beer, P. D., Kocian, O., Mortimer, R. J., and Ridgway, C., 1991, Syntheses, coordination, spectroscopy and electropolymerisation studies of new alkynyl and vinyl linked benzo-and aza-crown ether-bipyridyl ruthenium(II) complexes. Spectrochemical recognition of group IA/IIA metal cations, J. Chem. Soc., Chem. Commun. 1991: 1460–1463.CrossRefGoogle Scholar
  66. 65.
    Yoon, D. I., Berg-Brennan, C. A., Lu, H., and Hupp, J. T., 1992, Synthesis and preliminary photophysical studies of intramolecular electron transfer in crown-linked donor—(chromophore—) acceptor complexes, Inorg. Chem. 31: 3192–3194.Google Scholar
  67. 66.
    MacQueen, D. B., and Schanze, K. S., 1991, Cation-controlled photophysics in a Re(I) fluoroionophore, J. Am. Chem. Soc. 113: 6108–6110.CrossRefGoogle Scholar
  68. 67.
    Rawle, S. C., Moore, P., and Alcock, N. W., 1992, Synthesis and coordination chemistry of I-(2’,2“-bipyridyl-5’-yl-methyl)- 1,4,8,11-tetraazacyclotetradecane LII. Quenching of fluorescence from [Ru(bipy)2(L1)]2+ by coordination of Ni Z+ or Cut+ in the cyclam cavity, J. Chem. Soc., Chem. Commun. 1992: 684–687.CrossRefGoogle Scholar
  69. 68.
    Sun, H., and Hoffman, M. Z., 1993, Protonation of the excited states of ruthenium (II) complexes containing 2,2’-bipyridine, 2,2’bipyrazine and 2,2’-bipyrimidine ligands in aqueous solution, J. Phys. Chem. 97: 5014–5018.CrossRefGoogle Scholar
  70. 69.
    Park, J. W., Ahn, J., and Lee, C. 1995, Dependence of the photo-physical and photochemical properties of the photosensitizer tris(4,4’-dicarboxy-2,2’-bipyridine)ruthenium (II) on pH, J. Photochem. Photobiol., A: Chem. 86: 89–95.CrossRefGoogle Scholar
  71. 70.
    de Silva, A. P., Gunaratne, H. Q. N., and Lynch, P. L. M., 1995, Luminescence and charge transfer. Part 4. “On—off’ fluorescent PET (photoinduced electron transfer) sensors with pyridine receptors: 1,3-diaryl-5-pyridyl-4,5-dihydropyrazoles, J Chem. Soc., Perkin Trans. 2 1995: 685–690.Google Scholar
  72. 71.
    Walsh, M., Ryan, E. M., O’Kennedy, R., and Vos, J. G., 1996, The pH dependence of the emitting properties of ruthenium polypyridyl complexes bound to poly-L-lysine, J. Inorg. Biochem. 63: 215–221.CrossRefGoogle Scholar
  73. 72.
    Giordano, P. J., Bock, C. R., and Wrighton, M. S., 1978, Excited state proton transfer of ruthenium(II) complexes of 4,7-dihydroxy-1,10phenanthroline. Increased acidity in the excited state, J. Am. Chem. Soc. 100: 6960–6965.CrossRefGoogle Scholar
  74. 73.
    Grigg, R., and Norbert, W. D. J. A., 1992, Luminescent pH sensors based on di(2,2’-bipyridyl) (5,5’-diaminomethyl-2,2’-bipyridyl)-ruthenium(II) complexes, J. Chem. Soc., Chem. Commun. 1992: 1300–1302.CrossRefGoogle Scholar
  75. 74.
    Fantini, S., Franceschini, M. A., Fishkin, J. B., Barbieri, B., and Gratton, E., 1994, Quantitative determination of the absorption spectra of chromophores in strongly scattering media: A light-emitting diode based technique, Appl. Opt. 33: 5204–5213.CrossRefGoogle Scholar
  76. 75.
    Lippitsch, M. E., and Wolfbeis, O. S., 1988, Fiber-optics oxygen sensor with the fluorescence decay time as the information carrier, Anal. Chim. Acta 205: 1–6.CrossRefGoogle Scholar
  77. 76.
    Sipior, J., Carter, G. M., Lakowicz, J. R., and Rao, G., 1996, Single quantum well light emitting diodes demonstrated as excitation sources for nanosecond phase-modulation fluorescence lifetime measurements, Rev. Sci. Instrum. 67: 3795–3798.CrossRefGoogle Scholar
  78. 77.
    Castellano, E. N., and Lakowicz, J. R., 1998, A water-soluble luminescence oxygen sensor, Photochem. Photobiol. 67: 179–183.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1999

Authors and Affiliations

  • Joseph R. Lakowicz
    • 1
  1. 1.University of Maryland School of MedicineBaltimoreUSA

Personalised recommendations