Abstract
Fourier transform infrared (FTIR) spectroscopy is one of the widely used vibrational spectroscopic methods in protein structural analysis. The protein solution sample loaded in demountable CaF2 liquid cell presents a challenge and is limited to high concentrations. Some researchers attempted the simpler solid-film sampling method for the collection of protein FTIR spectra. In this study, the solid-film sampling FTIR method was studied in detail. The secondary structure components of some globular proteins were determined by this sampling method, and the results were consistent with those data determined by the traditional solution sampling FTIR method and X-ray crystallography, indicating that this sampling method is feasible and efficient for the structural characterization of proteins. Furthermore, much lower protein concentrations (~0.5 mg/mL) were needed to obtain high-quality FTIR spectra, which expands the application of FTIR spectroscopy to almost the same concentration range used for circular dichroism and fluorescence spectroscopy, making comparisons among three commonly used techniques possible in protein studies.
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References
Smith BC. Fundamentals of Fourier transform infrared spectroscopy. Boca Raton: CRC Press; 1996.
Surewicz WK, Mantsch HH. New insight into protein secondary structure from resolution-enhanced infrared spectra. Biochim Biophys Acta Protein Struct Mol Enzymol. 1988;952:115–30.
Zhao Y, Yang H, Meng K, Yu S. Probing the Ca 2+/CaM-induced secondary structural and conformational changes in calcineurin. Int J Biol Macromol. 2014;64:453–7.
Fu C, Zhang J, Zheng Y, Xu H, Yu S. Binding of calmodulin changes the calcineurin regulatory region to a less dynamic conformation. Int J Biol Macromol. 2015;79:235–9.
Jiang Y, Li C, Nguyen X, Muzammil S, Towers E, Gabrielson J, et al. Qualification of FTIR spectroscopic method for protein secondary structural analysis. J Pharm Sci. 2011;100(11):4631–41.
Goormaghtigh E, Ruysschaert J-M, Raussens V. Evaluation of the information content in infrared spectra for protein secondary structure determination. Biophys J. 2006;90(8):2946–57.
Parker FS. Applications of infrared, Raman, and resonance Raman spectroscopy in biochemistry. Springer Science & Business Media; 1983.
Byler DM, Susi H. Examination of the secondary structure of proteins by deconvolved FTIR spectra. Biopolymers. 1986;25(3):469–87.
Susi H. Infrared spectroscopy—conformation. Methods Enzymol. 1972;26:455–72.
Griffiths P, Durig J. Analytical applications of FT-IR to molecular and biological systems. Dordrecht: Reidel; 1980. 11
Susi H, Timasheff SN, Stevens L. Infrared spectra and protein conformations in aqueous solutions I. The amide I band in H2O and D2O solutions. J Biol Chem. 1967;242(23):5460–6.
Gorga JC, Dong A, Manning MC, Woody RW, Caughey WS, Strominger JL. Comparison of the secondary structures of human class I and class II major histocompatibility complex antigens by Fourier transform infrared and circular dichroism spectroscopy. Proc Natl Acad Sci India. 1989;86(7):2321–5.
Lee DC, Haris PI, Chapman D, Mitchell RC. Determination of protein secondary structure using factor analysis of infrared spectra. Biochemistry. 1990;29(39):9185–93.
Yamamoto T, Tasumi M. FT-IR studies on thermal denaturation processes of ribonucleases A and S in H2O and D2O solutions. J Mol Struct. 1991;242:235–44.
Kong J, Yu S. Fourier transform infrared spectroscopic analysis of protein secondary structures. Acta Biochim Biophys Sin. 2007;39(8):549–59.
Fu C, Yang H, Wang M, Xiong H, Yu S. Serum albumin adsorbed on Au nanoparticles: structural changes over time induced by S–Au interaction. Chem Commun. 2015;51(17):3634–6.
Wang M, Fu C, Liu X, Lin Z, Yang N, Yu S. Probing the mechanism of plasma protein adsorption on Au and Ag nanoparticles with FT-IR spectroscopy. Nano. 2015;7(37):15191–6.
Wu G, Gao Z, Dong A, Yu S. Calcium-induced changes in calmodulin structural dynamics and thermodynamics. Int J Biol Macromol. 2012;50(4):1011–7.
Dong A, Huang P, Caughey WS. Redox-dependent changes in. beta.-extended chain and turn structures of cytochrome c in water solution determined by second derivative amide I infrared spectra. Biochemistry. 1992;31(1):182–9.
Kal'Nin N, Ven'yaminov SY. Quantitative measurement of the IR spectra of water solutions. J Appl Chem. 1988;49(4):1028–32.
Yang H, Yang S, Kong J, Dong A, Yu S. Obtaining information about protein secondary structures in aqueous solution using Fourier transform IR spectroscopy. Nat Protoc. 2015;10(3):382–96.
Dong A, Randolph TW, Carpenter JF. Entrapping intermediates of thermal aggregation in alpha-helical proteins with low concentration of guanidine hydrochloride. J Biol Chem. 2000;275(36):27689–93.
Ismail AA, Mantsch HH, Wong PT. Aggregation of chymotrypsinogen: portrait by infrared spectroscopy. Biochim Biophys Acta Protein Struct Mol Enzymol. 1992;1121(1):183–8.
Fabian H, Mantsch HH. Ribonuclease A revisited: infrared spectroscopic evidence for lack of native-like secondary structures in the thermally denatured state. Biochemistry. 1995;34(41):13651–5.
Martínez A, Haavik J, Flatmark T, Arrondo JLR, Muga A. Conformational properties and stability of tyrosine hydroxylase studied by infrared spectroscopy EFFECT OF IRON/CATECHOLAMINE BINDING AND PHOSPHORYLATION. J Biol Chem. 1996;271(33):19737–42.
Lee TH, Cheng WT, Lin SY. Thermal stability and conformational structure of salmon calcitonin in the solid and liquid states. Biopolymers. 2010;93(2):200–7.
Seshadri S, Khurana R, Fink AL. Fourier transform infrared spectroscopy in analysis of protein deposits. Methods Enzymol. 1999;309:559–76.
Aiba H, Fujimoto S, Ozaki N. Molecular cloning and nucleotide sequencing of the gene for E. coli cAMP receptor protein. Nucleic Acids Res. 1982;10(4):1345–61.
Luthra S, Kalonia DS, Pikal MJ. Effect of hydration on the secondary structure of lyophilized proteins as measured by Fourier transform infrared (FTIR) spectroscopy. J Pharm Sci. 2007;96(11):2910–21.
Dong A, Caughey WS. Infrared methods for study of hemoglobin reactions and structures. Methods Enzymol. 1994;232:139–75.
Dong A, Huang P, Caughey WS. Protein secondary structures in water from second-derivative amide I infrared spectra. Biochemistry. 1990;29(13):3303–8.
Savitzky A, Golay MJ. Smoothing and differentiation of data by simplified least squares procedures. Anal Chem. 1964;36(8):1627–39.
Dong A, Prestrelski SJ, Allison SD, Carpenter JF. Infrared spectroscopic studies of lyophilization-and temperature-induced protein aggregation. J Pharm Sci. 1995;84(4):415–24.
Levitt M, Greer J. Automatic identification of secondary structure in globular proteins. J Mol Biol. 1977;114(2):181–239.
Richardson JS. The anatomy and taxonomy of protein structure. Adv Protein Chem. 1981;34:167–339.
Jackson M, Mantsch HH. The use and misuse of FTIR spectroscopy in the determination of protein structure. Crit Rev Biochem Mol. 1995;30(2):95–120.
Uversky VN, Permiakov EAe. Methods in protein structure and stability analysis: luminescence spectroscopy and circular dichroism. Nova Publishers; 2007.
Dong A, Jones LS, Kerwin BA, Krishnan S, Carpenter JF. Secondary structures of proteins adsorbed onto aluminum hydroxide: infrared spectroscopic analysis of proteins from low solution concentrations. Anal Biochem. 2006;351(2):282–9.
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This work was supported by the National Natural Science Foundation of China (Nos. 31470786 and 21275032).
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Zhang, J., Zhang, X., Zhang, F. et al. Solid-film sampling method for the determination of protein secondary structure by Fourier transform infrared spectroscopy. Anal Bioanal Chem 409, 4459–4465 (2017). https://doi.org/10.1007/s00216-017-0390-y
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DOI: https://doi.org/10.1007/s00216-017-0390-y