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| Time-Resolved Fluorescence and Thermodynamic Properties of Staphylococcal Nuclease |
| CHANG Meng-fang1, JIA Meng-hui2, LI Lei1, CHEN Jin-quan1, XU Jian-hua1* |
1. State Key Laboratory of Precision Spectroscopy, East China Normal University, Shanghai 200062, China
2. Shanghai Institute of Optics and Fine Mechanics, Chinese Academy of Sciences, Shanghai 201800, China |
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Abstract Staphylococcal nuclease is a small globular protein, whose variants are widely used in the researches on protein folding. Different from methods and techniques reported in published papers, fluorescence dynamics of tryptophan residues in two staphylococcal nuclease (SNase) variants Δ+PHS and Δ+PHS+I92A were investigated by time-correlated single photon counting (TCSPC) and femtosecond fluorescence upconversion techniques, combined with UV absorption and steady-state fluorescence spectroscopy. Based on the analysis, structures and thermal stabilities of the two SNase variants were studied. The results proved that tryptophan could be used as an endogenous probe for the structural folding and thermal stability of the SNase variants. Decay associated spectra (DAS) of SNase variants showed different changing trends upon temperatures. According to this, structural folding and thermal stability of the two variants were analyzed. Time-resolved emission spectra (TRES) demonstrated the 0.5 ns continuous spectral relaxation process of tryptophan residue, in which the spectral shift showed the compactness difference of folding structures of the two SNase variants. In femtosecond upconversion results, DAS of 0.5 ps lifetime had “positive blue edge and negative red edge”, which showed relaxation effects on tryptophan residues in SNase variants. Moreover, the lifetime of 200 ps indicated the electron transfer between tryptophan residues and surrounding quenching group. Analysis of time-resolved anisotropy showed that the tryptophan residues had independent segmental motion in the protein system, and its intensity was related to the thermal stability of SNase variants and the overall effect of thermal motion. Time-resolved fluorescence measurement and analysis of tryptophan residue helped to investigate the relationship between structure and function of protein.
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Received: 2017-04-20
Accepted: 2017-09-02
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Corresponding Authors:
XU Jian-hua
E-mail: jhxu@phy.ecnu.edu.cn
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[1] HUANG Ke-han, QIN Cui-fang, CAO Xiao-dan, et al(黄科翰，秦翠芳，曹潇丹，等). Spectroscopy and Spectral Analysis(光谱学与光谱分析), 2016, 36(12): 3973.
[2] QIN Cui-fang, LI Lei, YU Xian-tong, et al(秦翠芳，李 磊，俞宪同，等). Spectroscopy and Spectral Analysis(光谱学与光谱分析), 2017, 37(2): 476.
[3] Chang M, Li L, Cao X, et al. Acta Physico-Chimica Sinica(物理化学学报), 2017, 33(5): 1065.
[4] Frauenfelder H, Sligar S G, Wolynes P G. Science, 1991, 254(5038): 1598.
[5] Xu J, Toptygin D, Graver K J, et al. Journal of the American Chemical Society, 2006, 128(4): 1214.
[6] Roche J, Dellarole M, Caro J A, et al. Biochemistry, 2012, 51(47): 9535.
[7] Roche J, Caro J A, Dellarole M, et al. Proteins, 2013, 81(6): 1069.
[8] Roche J, Dellarole M, Caro J A, et al. Journal of the American Chemical Society, 2013, 135(39): 14610.
[9] Roche J, Caro J A, Norberto D R, et al. Proceedings of the National Academy of Sciences of the United States of America, 2012, 109(18): 6945.
[10] Jia M, Yang J, Qin Y, et al. Journal of Physical Chemistry Letters, 2015, 6(24): 5100.
[11] Jia M, Yi H, Chang M, et al. Journal of Photochemistry and Photobiology B: Biology, 2015, 149: 243.
[12] Lakowicz J R. Principles of Fluorescence Spectroscopy. 3rd ed. New York: Springer Science & Business Media, 2006.
[13] Zhong D. Advances in Chemical Physics, 2009, 143: 83.
[14] Isom D G, Cannon B R, Casta X, et al. Proceedings of the National Academy of Sciences of the United States of America, 2008, 105(46): 17784.
[15] Royer C, Winter R. Current Opinion in Colloid & Interface Science, 2011, 16(6): 568.
[16] Petrich J W, Chang M C, McDonald D B, et al. Journal of the American Chemical Society, 1983, 105(12): 3824. |
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