Single-Molecule Spectroscopy of Unfolded Proteins

Instrumental Analysis of Intrinsically Disordered Proteins: Assessing Structure and Conformation

Volumn , Issue , 2010, Pages 369-389

  Schuler, Benjamin ^a  

^a UNIVERSITY OF ZURICH   (Switzerland)

Author keywords

New protein labeling techniques and fluorophore development instrumentation, data acquisition, data analysis and theory; Single molecule spectroscopy of unfolded state; Single molecule techniques for unfolded proteins

Indexed keywords

EID: 84885492529     PISSN: None     EISSN: None     Source Type: Book    
DOI: 10.1002/9780470602614.ch13     Document Type: Chapter

References (112)

1. Antonik, M., S. Felekyan, A. Gaiduk, and C. A. M. Seidel. 2006. Separating structural heterogeneities from stochastic variations in fluorescence resonance energy transfer distributions via photon distribution analysis. J Phys Chem B 110: 6970-8.
2. Axelrod, D., T. P. Burghardt, and N. L. Thompson. 1984. Total internal-reflection fluorescence. Annu Rev Biophys Bioeng 13: 247-68.
3. Baldini, G., F. Cannone, and G. Chirico. 2005. Pre-unfolding resonant oscillations of single green fluorescent protein molecules. Science 309: 1096-100.
4. Best, R., K. Merchant, I. V. Gopich, B. Schuler, A. Bax, and W. A. Eaton. 2007. Effect of flexibility and cis residues in single molecule FRET studies of polyproline. Proc Natl Acad. Sci U S A 104: 18964-9.
5. Best, R. B., and G. Hummer. 2006. Diffusive model of protein folding dynamics with Kramers turnover in rate. Phys Rev Lett96: 228104.
6. Böhmer, M., and J. Enderlein. 2003. Fluorescence spectroscopy of single molecules under ambient conditions: methodology and technology. Chem Phys Chem 4: 793-808.
7. Borgia, A., P. M. Williams, and J. Clarke. 2008. Single-molecule studies of protein folding. Annu Rev Biochem 77: 101-25.
8. Boukobza, E., A. Sonnenfeld, and G. Haran. 2001. Immobilization in surface-tethered lipid vesicles as a new tool for single biomolecule spectroscopy. J Phys Chem B 105: 12165-70.
9. Bryngelson, J. D., J. N. Onuchic, N. D. Socci, and P. G. Wolynes. 1995. Funnels, pathways, and the energy landscape of protein folding: a synthesis. Proteins 21: 167-95.
10. Buscaglia, M., L. J. Lapidus, W. A. Eaton, and J. Hofrichter. 2006. Effects of denaturants on the dynamics of loop formation in polypeptides. Biophys J 91: 276-88.
11. Bustamante, C., Y. R. Chemla, N. R. Forde, and D. Izhaky. 2004. Mechanical processes in biochemistry. Annu Rev Biochem 73: 705-48.
12. Cannone, F., S. Bologna, B. Campanini, A. Diaspro, S. Bettati, A. Mozzarelli, and G. Chirico. 2005. Tracking unfolding and refolding of single GFPmut2 molecules. Biophysl J 89: 2033-45.
13. Chahine, J., R. J. Oliveira, V. B. P. Leite, and J. Wang. 2007. Configuration-dependent diffusion can shift the kinetic transition state and barrier height of protein folding. Proc Natl Acad Sci U S A 104: 14646-51. 384 INSTRUMENTAL ANALYSIS OF INTRINSICALLY DISORDERED PROTEINS
14. Chan, W. C., D. J. Maxwell, X. Gao, R. E. Bailey, M. Han, and S. Nie. 2002. Luminescent quantum dots for multiplexed biological detection and imaging. Curr Opin Biotechnol 13: 40-6.
15. Chattopadhyay, K., E. L. Elson, and C. Frieden. 2005. The kinetics of conformational fluctuations in an unfolded protein measured by fluorescence methods. Proc Natl Acad Sci U S A 102: 2385-9.
16. Chattopadhyay, K., S. Saffarian, E. L. Elson, and C. Frieden. 2005. Measuring unfolding of proteins in the presence of denaturant using fluorescence correlation spectroscopy. Biophys J 88: 1413-22.
17. Clamme, J. -P., and A. A. Deniz. 2005. Three-color single-molecule fluorescence resonance energy transfer. Chem Phys Chem 6: 74-7.
18. Cropp, T. A., and P. G. Schultz. 2004. An expanding genetic code. Trends Genet 20: 625-30.
19. David, R., M. P. Richter, and A. G. Beck-Sickinger. 2004. Expressed protein ligation. Method and applications. Eur J Biochem 271: 663-77.
20. Dawson, P. E., and S. B. Kent. 2000. Synthesis of native proteins by chemical ligation. Annu Rev Biochem 69: 923-60.
21. Deniz, A. A., M. Dahan, J. R. Grunwell, T. J. Ha, A. E. Faulhaber, D. S. Chemla, S. Weiss, and P. G. Schultz. 1999. Single-pair fluorescence resonance energy transfer on freely diffusing molecules: observation of Forster distance dependence and subpopulations. Proc Natl Acad Sci U S A 96: 3670-5.
22. Deniz, A. A., T. A. Laurence, G. S. Beligere, M. Dahan, A. B. Martin, D. S. Chemla, P. E. Dawson, P. G. Schultz, and S. Weiss. 2000. Single-molecule protein folding: diffusion fluorescence resonance energy transfer studies of the denaturation of chymotrypsin inhibitor 2. Proc Natl Acad Sci U S A 97: 5179-84.
23. Deniz, A. A., T. A. Laurence, M. Dahan, D. S. Chemla, P. G. Schultz, and S. Weiss. 2001. Ratiometric single-molecule studies of freely diffusing biomolecules. Annu Rev Phys Chem 52: 233-53.
24. Dyson, H. J., and P. E. Wright. 2005. Intrinsically unstructured proteins and their functions. Nat Rev Mol Cell Biol 6: 197-208.
25. Dyson, H. J., and P. E. Wright. 2004. Unfolded proteins and protein folding studied by NMR. Chem Rev 104: 3607-22.
26. Eigen, M., and R. Rigler. 1994. Sorting single molecules: application to diagnostics and evolutionary biotechnology. Proc Natl Acad Sci U S A 91: 5740-7.
27. Fisher, T. E., A. F. Oberhauser, M. Carrion-Vazquez, P. E. Marszalek, and J. M. Fernandez. 1999. The study of protein mechanics with the atomic force microscope. Trends Biochem Sci 24: 379-84.
28. Forman, J. R., and J. Clarke. 2007. Mechanical unfolding of proteins: insights into biology, structure and folding. Curr Opin Struct Biol 17: 58-66.
29. Förster, T. 1948. Zwischenmolekulare Energiewanderung und Fluoreszenz. Annalen der Physik 6: 55-75.
30. Gopich, I. V., and A. Szabo. 2005. Theory of photon statistics in single-molecule F ö rster resonance energy transfer. J Chem Phys 122: 1-18.
31. Gopich, I. V., and A. Szabo. 2006. Theory of the statistics of kinetic transitions with application to single-molecule enzyme catalysis. J Chem Phys 124: 154712. SINGLE-MOLECULE SPECTROSCOPY OF UNFOLDED PROTEINS 385
32. Grinvald, A., E. Haas, and I. Z. Steinber. 1972. Evaluation of distribution of distances between energy donors and acceptors by fluorescence decay. Proc Natl Acad Sci U S A 69: 2273-2277.
33. Groll, J., E. V. Amirgoulova, T. Ameringer, C. D. Heyes, C. Rocker, G. U. Nienhaus, and M. Moller. 2004. Biofunctionalized, ultrathin coatings of cross-linked star-shaped poly(ethylene oxide) allow reversible folding of immobilized proteins. J Am Chem Soc 126: 4234-9.
34. Ha, T. 2001. Single-molecule fluorescence resonance energy transfer. Methods 25: 78-86.
35. Ha, T., T. Enderle, D. F. Ogletree, D. S. Chemla, P. R. Selvin, and S. Weiss. 1996. Probing the interaction between two single molecules: fluorescence resonance energy transfer between a single donor and a single acceptor. Proc Natl Acad Sci U S A 93: 6264-8.
36. Haas, E., E. Katchalskikatzir, and I. Z. Steinberg. 1978. Brownian-motion of ends of oligopeptide chains in solution as estimated by energy-transfer between chain ends. Biopolymers 17: 11-31.
37. Haas, E., E. Katchalskikatzir, and I. Z. Steinberg. 1978. Brownian-motion of ends of oligopeptide chains in solution as estimated by energy-transfer between chain ends. Biopolymers 17: 11-31.
38. Hess, S. T., S. Huang, A. A. Heikal, and W. W. Webb. 2002. Biological and chemical applications of fluorescence correlation spectroscopy: a review. Biochemistry 41: 697-705.
39. Heyduk, T. 2002. Measuring protein conformational changes by FRET/LRET. Curr Opin Biotechnol 13: 292-6.
40. Hillger, F., D. Hänni, D. Nettels, S. Geister, M. Grandin, M. Textor, and B. Schuler. 2008. Probing protein-chaperone interactions with single molecule fluorescence spectroscopy. Angew Chem Int Ed Engl 47: 6184-8.
41. Hillger, F., D. Nettels, S. Dorsch, and B. Schuler. 2007. Detection and analysis of protein aggregation with confocal single molecule fluorescence spectroscopy. J Fluoresc 17: 759-65.
42. Hoffmann, A., A. Kane, D. Nettels, D. E. Hertzog, P. Baumgartel, J. Lengefeld, G. Reichardt, D. A. Horsley, R. Seckler, O. Bakajin, and B. Schuler. 2007. Mapping protein collapse with single-molecule fluorescence and kinetic synchrotron radiation circular dichroism spectroscopy. Proc Natl Acad Sci U S A 104: 105-10.
43. Hohng, S., C. Joo, and T. Ha. 2004. Single-molecule three-color FRET. Biophys J 87: 1328-37.
44. Huang, F., S. Sato, T. D. Sharpe, L. M. Ying, and A. R. Fersht. 2007. Distinguishing between cooperative and unimodal downhill protein folding. Proc Natl Acad Sci U S A 104: 123-7.
45. Kapanidis, A. N., N. K. Lee, T. A. Laurence, S. Doose, E. Margeat, and S. Weiss. 2004. Fluorescence-aided molecule sorting: analysis of structure and interactions by alternating-laser excitation of single molecules. Proc Natl Acad Sci U S A 101: 8936-41.
46. Kapanidis, A. N., and S. Weiss. 2002. Fluorescent probes and bioconjugation chemistries for single-molecule fluorescence analysis of biomolecules. J Chem Phys 117: 10953-64. 386 INSTRUMENTAL ANALYSIS OF INTRINSICALLY DISORDERED PROTEINS
47. Kinoshita, M., K. Kamagata, A. Maeda, Y. Goto, T. Komatsuzaki, and S. Takahashi. 2007. Development of a technique for the investigation of folding dynamics of single proteins for extended time periods. Proc Natl Acad Sci U S A 104: 10453-58.
48. Kohn, J. E., I. S. Millett, J. Jacob, B. Zagrovic, T. M. Dillon, N. Cingel, R. S. Dothager, S. Seifert, P. Thiyagarajan, T. R. Sosnick, M. Z. Hasan, V. S. Pande, I. Ruczinski, S. Doniach, and K. W. Plaxco. 2004. Random-coil behavior and the dimensions of chemically unfolded proteins. Proc Natl Acad Sci U S A 101: 12491-6.
49. Kremer, W., B. Schuler, S. Harrieder, M. Geyer, W. Gronwald, C. Welker, R. Jaenicke, and H. R. Kalbitzer. 2001. Solution NMR structure of the cold-shock protein from the hyperthermophilic bacterium Thermotoga maritima. Eur J Biochem 268: 2527-39.
50. Kubelka, J., J. Hofrichter, and W. A. Eaton. 2004. The protein folding " speed limit ". Curr Opin Struct Biol 14: 76-88.
51. Kuzmenkina, E. V., C. D. Heyes, and G. U. Nienhaus. 2005. Single-molecule Forster resonance energy transfer study of protein dynamics under denaturing conditions. Proc Natl Acad Sci U S A 102: 15471-6.
52. Kuzmenkina, E. V., C. D. Heyes, and G. U. Nienhaus. 2006. Single-molecule FRET study of denaturant induced unfolding of RNase H. J Mol Biol 357: 313-24.
53. Kuzmenkina, E. V., C. D. Heyes, and G. U. Nienhaus. 2006. Single-molecule FRET study of denaturant induced unfolding of RNase H. J Mol Biol 357: 313-24.
54. Laurence, T. A., X. X. Kong, M. Jager, and S. Weiss. 2005. Probing structural heterogeneities and fluctuations of nucleic acids and denatured proteins. Proc Natl Acad Sci U S A 102: 17348-53.
55. Lipman, E. A., B. Schuler, O. Bakajin, and W. A. Eaton. 2003. Single-molecule measurement of protein folding kinetics. Science 301: 1233-5.
56. Lippitz, M., W. Erker, H. Decker, K. E. van Holde, and T. Basche. 2002. Two-photon excitation microscopy of tryptophan-containing proteins. Proc Natl Acad Sci U S A 99: 2772-7.
57. Magg, C., J. Kubelka, G. Holtermann, E. Haas, and F. X. Schmid. 2006. Specificity of the initial collapse in the folding of the cold shock protein. J Mol Biol 360: 1067-80.
58. Magg, C., and F. X. Schmid. 2004. Rapid collapse precedes the fast two-state folding of the cold shock protein. J Mol Biol 335: 1309-23.
59. McCarney, E. R., J. H. Werner, S. L. Bernstein, I. Ruczinski, D. E. Makarov, P. M. Goodwin, and K. W. Plaxco. 2005. Site-specific dimensions across a highly denatured protein; a single molecule study. J Mol Biol 352: 672-82.
60. Merchant, K. A., R. B. Best, J. M. Louis, I. V. Gopich, and W. A. Eaton. 2007. Characterizing the unfolded states of proteins using single-molecule FRET spectroscopy and molecular simulations. Proc Natl Acad Sci U S A 104: 1528-33.
61. Michalet, X., A. N. Kapanidis, T. Laurence, F. Pinaud, S. Doose, M. Pflughoefft, and S. Weiss. 2003. The power and prospects of fluorescence microscopies and spectroscopies. Annu Rev Biophys Biomol Struct 32: 161-82. SINGLE-MOLECULE SPECTROSCOPY OF UNFOLDED PROTEINS 387
62. Michalet, X., S. Weiss, and M. Jäger. 2006. Single-molecule fluorescence studies of protein folding and conformational dynamics. Chem Rev 106: 1785-813.
63. Millett, I. S., S. Doniach, and K. W. Plaxco. 2002. Toward a taxonomy of the denatured state: small angle scattering studies of unfolded proteins. Adv Protein Chem 62: 241-62.
64. Millet, I. S., L. E. Townsley, F. Chiti, S. Doniach, and K. W. Plaxco. 2002. Equilibrium collapse and the kinetic " foldability " of proteins. Biochemistry 41: 321-5.
65. Moerner, W. E. 2002. A dozen years of single-molecule spectroscopy in physics, chemistry, and biophysics. J Phys Chem B 106: 910-27.
66. Möglich, A., K. Joder, and T. Kiefhaber. 2006. End-to-end distance distributions and intrachain diffusion constants in unfolded polypeptide chains indicate intramolecular hydrogen bond formation. Proc Natl Acad Sci U S A 103: 12394-9.
67. Mujumdar, R. B., L. A. Ernst, S. R. Mujumdar, C. J. Lewis, and A. S. Waggoner. 1993. Cyanine dye labeling reagents: sulfoindocyanine succinimidyl esters. Bioconjug Chem 4: 105-11.
68. Mukhopadhyay, S., R. Krishnan, E. A. Lemke, S. Lindquist, and A. A. Deniz. 2007. A natively unfolded yeast prion monomer adopts an ensemble of collapsed and rapidly fluctuating structures. Proc Natl Acad Sci U S A 104: 2649-54.
69. Müller, B. K., E. Zaychikov, C. Br äuchle, and D. C. Lamb. 2005. Pulsed interleaved excitation. Biophys J 89: 3508-22.
70. Müller, B. K., E. Zaychikov, C. Br äuchle, and D. C. Lamb. 2005. Pulsed interleaved excitation. Biophys J 89: 3508-22.
71. Nettels, D., I. V. Gopich, A. Hoffmann, and B. Schuler. 2007. Ultrafast dynamics of protein collapse from single-molecule photon statistics. Proc Natl Acad Sci U S A 104: 2655-60.
72. Nettels, D., A. Hoffmann, and B. Schuler. 2008. Unfolded protein and peptide dynamics investigated with single-molecule FRET and correlation spectroscopy from picoseconds to seconds. J Phys Chem B 112: 6137-46.
73. Nettels, D., and B. Schuler. 2007. Subpopulation-resolved photon statistics of single-molecule energy transfer dynamics. IEEE J Sel Top Quant Electron 13: 990-5.
74. Neuweiler, H., S. Doose, and M. Sauer. 2005. A microscopic view of miniprotein folding: enhanced folding efficiency through formation of an intermediate. Proc Natl Acad Sci U S A 102: 16650-5.
75. Neuweiler, H., M. Lollmann, S. Doose, and M. Sauer. 2007. Dynamics of unfolded polypeptide chains in crowded environment studied by fluorescence correlation spectroscopy. J Mol Biol 365: 856-69.
76. Nienhaus, G. U. 2006. Exploring protein structure and dynamics under denaturing conditions by single-molecule FRET analysis. Macromol Biosci 6: 907-22.
77. Nir, E., X. Michalet, K. M. Hamadani, T. A. Laurence, D. Neuhauser, Y. Kovchegov, and S. Weiss. 2006. Shot-noise limited single-molecule FRET histograms: comparison between theory and experiments. J Phys Chem B 110: 22103-24.
78. O'Brien, E. P., G. Ziv, G. Haran, B. R. Brooks, and D. Thirumalai. 2008. Effects of denaturants and osmolytes on proteins are accurately predicted by the molecular transfer model. Proc Natl Acad Sci U S A 105: 13403-8. 388 INSTRUMENTAL ANALYSIS OF INTRINSICALLY DISORDERED PROTEINS
79. Panchuk-Voloshina, N., R. P. Haugland, J. Bishop-Stewart, M. K. Bhalgat, P. J. Millard, F. Mao, and W. Y. Leung. 1999. Alexa dyes, a series of new fluorescent dyes that yield exceptionally bright, photostable conjugates. J Histochem Cytochem 47: 1179-88.
80. Plaxco, K. W., I. S. Millett, D. J. Segel, S. Doniach, and D. Baker. 1999. Chain collapse can occur concomitantly with the rate-limiting step in protein folding. Nat Struct Biol 6: 554-6.
81. Ratner, V., E. Kahana, M. Eichler, and E. Haas. 2002. A general strategy for site-specific double labeling of globular proteins for kinetic FRET studies. Bioconjug Chem 13: 1163-70.
82. Rhoades, E., M. Cohen, B. Schuler, and G. Haran. 2004. Two-state folding observed in individual protein molecules. J Am Chem Soc 126: 14686-7.
83. Rhoades, E., E. Gussakovsky, and G. Haran. 2003. Watching proteins fold one molecule at a time. Proc Natl Acad Sci U S A 100: 3197-202.
84. Sako, Y., and T. Uyemura. 2002. Total internal reflection fluorescence microscopy for single-molecule imaging in living cells. Cell Struct Funct 27: 357-65.
85. Schuler, B. 2007. Application of single molecule Forster resonance energy transfer to protein folding. Methods Mol Biol 350: 115-38.
86. Schuler, B. 2007. Application of single molecule Forster resonance energy transfer to protein folding. Methods Mol Biol 350: 115-38.
87. Schuler, B. 2005. Single-molecule fluorescence spectroscopy of protein folding. Chemphyschem 6: 1206-20.
88. Schuler, B., and W. A. Eaton. 2008. Protein folding studied by single-molecule FRET. Curr Opin Struct Biol 18: 16-26.
89. Schuler, B., E. A. Lipman, and W. A. Eaton. 2002. Probing the free-energy surface for protein folding with single-molecule fluorescence spectroscopy. Nature 419: 743-7.
90. Schuler, B., E. A. Lipman, P. J. Steinbach, M. Kumke, and W. A. Eaton. 2005. Polyproline and the " spectroscopic ruler " revisited with single molecule fluorescence. Proc Natl Acad Sci U S A 102: 2754-9.
91. Schuler, B., and L. K. Pannell. 2002. Specific labeling of polypeptides at amino-terminal cysteine residues using Cy5-benzyl thioester. Bioconjug Chem 13: 1039-43.
92. Selvin, P. R. 2000. The renaissance of fluorescence resonance energy transfer. Nature Struct Biol 7: 730-4.
93. Sharma, S., K. Chakraborty, B. K. Muller, N. Astola, Y. C. Tang, D. C. Lamb, M. Hayer-Hartl, and F. U. Hartl. 2008. Monitoring protein conformation along the pathway of chaperonin-assisted folding. Cell 133: 142-53.
94. Sherman, E., and G. Haran. 2006. Coil-globule transition in the denatured state of a small protein. Proc Natl Acad Sci U S A 103: 11539-43.
95. Shimomura, O. 2005. The discovery of aequorin and green fluorescent protein. J Microsc 217: 1-15.
96. Stryer, L. 1978. Fluorescence energy transfer as a spectroscopic ruler. Annu Rev Biochem 47: 819-46. SINGLE-MOLECULE SPECTROSCOPY OF UNFOLDED PROTEINS 389
97. Stryer, L., and R. P. Haugland. 1967. Energy transfer: a spectroscopic ruler. Proc Natl Acad Sci U S A 58: 719-26.
98. Taguchi, H., T. Ueno, H. Tadakuma, M. Yoshida, and T. Funatsu. 2001. Single-molecule observation of protein-protein interactions in the chaperonin system. Nat Biotechnol 19: 861-5.
99. Teilum, K., F. M. Poulsen, and M. Akke. 2006. The inverted chevron plot measured by NMR relaxation reveals a native-like unfolding intermediate in acyl-CoA binding protein. Proc Natl Acad Sci U S A 103: 6877-82.
100. Teilum, K., F. M. Poulsen, and M. Akke. 2006. The inverted chevron plot measured by NMR relaxation reveals a native-like unfolding intermediate in acyl-CoA binding protein. Proc Natl Acad Sci U S A 103: 6877-82.
101. Ueno, T., H. Taguchi, H. Tadakuma, M. Yoshida, and T. Funatsu. 2004. GroEL mediates protein folding with a two successive timer mechanism. Mol. Cell 14: 423-34.
102. Uversky, V. N., C. J. Oldfield, and A. K. Dunker. 2005. Showing your ID: intrinsic disorder as an ID for recognition, regulation and cell signaling. J Mol Recognit 18: 343-84.
103. Uversky, V. N., C. J. Oldfield, and A. K. Dunker. 2005. Showing your ID: intrinsic disorder as an ID for recognition, regulation and cell signaling. J Mol Recognit 18: 343-84.
104. Wahl, M., F. Koberling, M. Patting, H. Rahn, and R. Erdmann. 2004. Time-resolved confocal fluorescence imaging and spectrocopy system with single molecule sensitivity and sub-micrometer resolution. Curr Pharm Biotechnol 5: 299-308.
105. Wang, Z. S., and D. E. Makarov. 2003. Nanosecond dynamics of single polypeptide molecules revealed by photoemission statistics of fluorescence resonance energy transfer: a theoretical study. J Phys Chem B 107: 5617-22.
106. Weiss, S. 1999. Fluorescence spectroscopy of single biomolecules. Science 283: 1676-83.
107. White, S. S., S. Balasubramanian, D. Klenerman, and L. M. Ying. 2006. A simple nanomixer for single-molecule kinetics measurements. Angew Chem Int Ed Engl 45: 7540-3.
108. Widengren, J., V. Kudryavtsev, M. Antonik, S. Berger, M. Gerken, and C. A. M. Seidel. 2006. Single-molecule detection and identification of multiple species by multiparameter fluorescence detection. Anal Chem 78: 2039-50.
109. Yamaguchi, J., N. Nemoto, T. Sasaki, A. Tokumasu, Y. Mimori-Kiyosue, T. Yagi, and T. Funatsu. 2001. Rapid functional analysis of protein-protein interactions by fluorescent C-terminal labeling and single-molecule imaging. FEBS Lett502: 79-83.
110. Yamasaki, R., M. Hoshino, T. Wazawa, Y. Ishii, T. Yanagida, Y. Kawata, T. Higurashi, K. Sakai, J. Nagai, and Y. Goto. 1999. Single molecular observation of the interaction of GroEL with substrate proteins. J Mol Biol 292: 965-72.
111. Zhuang, X., and M. Rief. 2003. Single-molecule folding. Curr Opin Struct Biol 13: 88-97.
112. Zondervan, R., F. Kulzer, H. van der Meer, J. A. Disselhorst, and M. Orrit. 2006. Laser-driven microsecond temperature cycles analyzed by fluorescence polarization microscopy. Biophys J 90: 2958-69.