Abstract
Solution studies permit a direct investigation of the particles on a well-defined environment. Fluorescence, circular dichroism, scattering, and calorimetry provide, individually, very important information among the protein structure, overall shape, and thermodynamic equilibrium. In this work, a combination of these techniques is presented for the study of denaturation induced by temperature of two well-known proteins, Henn Egg lysozyme and bovine serum albumin. A detailed thermodynamic and structural investigation is shown for these proteins, providing interesting information on the thermal-induced changes in the protein structure and aggregation behavior.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
D. Voet, J.G. Voet, C.W. Pratt, Fundamentals of Biochemistry: Life at the Molecular Level, 2nd edn. (Wiley, New York, 2005)
D. Otzen, Protein-surfactant interactions: a tale of many states. Biochimica Et Biophysica Acta-Proteins and Proteomics 1814(5), 562–591 (2011)
M. Selmer, C.M. Dunham, F.V. Murphy, A. Weixlbaumer, S. Petry, A.C. Kelley, J.R. Weir, V. Ramakrishnan, Structure of the 70S ribosome complexed with mRNA and tRNA. Science 313(5795), 1935–1942 (2006)
L.J. Worrall, C. Hong, M. Vuckovic, W. Deng, J.R.C. Bergeron, D.D. Majewski, R.K. Huang, T. Spreter, B.B. Finlay, Z. Yu, N.C.J. Strynadka, Near-atomic-resolution cryo-EM analysis of the Salmonella T3S injectisome basal body. Nature 540(7634), 597-+ (2016)
C.L.P. Oliveira, in Current Trends in X-ray Crystallography, ed. by D. A. Chandrasekaran. Investigating macromolecular complexes in solution by small angle X-ray scattering (InTech, Rijeka, 2011), pp. 367–392
D.I. Svergun, M.H.J. Koch, P.A. Timmins, R.P. May, Small Angle X-ray and Neutron Scattering from Solutions of Biological Macromolecules (Oxford University Press, Oxford, 2013)
A. Lobley, L. Whitmore, B.A. Wallace, DICHROWEB: an interactive website for the analysis of protein secondary structure from circular dichroism spectra. Bioinformatics 18(1), 211–212 (2002)
L. Whitmore, B.A. Wallace, DICHROWEB, an online server for protein secondary structure analyses from circular dichroism spectroscopic data. Nucleic Acids Res. 32, W668–W673 (2004)
L. Whitmore, B.A. Wallace, Protein secondary structure analyses from circular dichroism spectroscopy: methods and reference databases. Biopolymers 89(5), 392–400 (2008)
A.V. Semenyuk, D.I. Svergun, GNOM—a program package for small-angle scattering data-processing. J. Appl. Crystallogr. 24, 537–540 (1991)
D.I. Svergun, Restoring low resolution structure of biological macromolecules from solution scattering using simulated annealing. Biophys. J. 76(6), 2879–2886 (1999)
M.V. Petoukhov, D. Franke, A.V. Shkumatov, G. Tria, A.G. Kikhney, M. Gajda, C. Gorba, H.D.T. Mertens, P.V. Konarev, D.I. Svergun, New developments in the ATSAS program package for small-angle scattering data analysis. J. Appl. Crystallogr. 45, 342–350 (2012)
M. Hirai, S. Arai, H. Iwase, T. Takizawa, Small-angle X-ray scattering and calorimetric studies of thermal conformational change of lysozyme depending on pH. J. Phys. Chem. B 102(7), 1308–1313 (1998)
Y. Sun, P.L.O. Filho, J.C. Bozelli Jr., J. Carvalho, S. Schreier, C.L.P. Oliveira, Unfolding and folding pathway of lysozyme induced by sodium dodecyl sulfate. Soft Matter 11(39), 7769–7777 (2015)
N.L. Almeida, C.L.P. Oliveira, I.L. Torriani, W. Loh, Calorimetric and structural investigation of the interaction of lysozyme and bovine serum albumin with poly(ethylene oxide) and its copolymers. Colloids and Surfaces B-Biointerfaces 38(1–2), 67–76 (2004)
V.V. Volkov, D.I. Svergun, Uniqueness of ab initio shape determination in small-angle scattering. J. Appl. Crystallogr. 36, 860–864 (2003)
C. Giancola, C. DeSena, D. Fessas, G. Graziano, G. Barone, DSC studies on bovine serum albumin denaturation—effects of ionic strength and SDS concentration. Int. J. Biol. Macromol. 20(3), 193–204 (1997)
K.A. Majorek, P.J. Porebski, A. Dayal, M.D. Zimmerman, K. Jablonska, A.J. Stewart, M. Chruszcz, W. Minor, Structural and immunologic characterization of bovine, horse, and rabbit serum albumins. Mol. Immunol. 52(3–4), 174–182 (2012)
K. Takeda, A. Wada, K. Yamamoto, Y. Moriyama, K. Aoki, Conformational change of bovine serum-albumin by heat-treatment. J. Protein Chem. 8(5), 653–659 (1989)
Borzova, V. A.; Markossian, K. A.; Chebotareva, N. A.; Kleymenov, S. Y.; Poliansky, N. B.; Muranov, K. O.; Stein-Margolina, V. A.; Shubin, V. V.; Markov, D. I.; Kurganov, B. I., Kinetics of thermal denaturation and aggregation of bovine serum albumin. Plos One, 11(4), 1–21 (2016)
Acknowledgments
The authors are grateful to Dra. Renata N. Bicev for the support on sample preparation and SAXS measurements, Dra. Laura Farkuh for the support on DSC measurements, Dr. Yang Sun for the support on fluorescence measurements and valuable discussions, and Dra. Valquiria P. Souza for the support on CD measurements. ASP is supported by CAPES and INCT-FCx. CLPO is supported by FAPESP, CNPQ, and INCT-FCx.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Perez, A.S., Oliveira, C.L.P. Thermal-Induced Denaturation and Aggregation Behavior of Lysozyme and Bovine Serum Albumin: a Thermodynamic and Structural Study. Braz J Phys 47, 524–531 (2017). https://doi.org/10.1007/s13538-017-0520-1
Received:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13538-017-0520-1