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High-sensitivity modulation differential scanning calorimetry of protein denaturation

Part 1. Two-state kinetics of thermal denaturation of Kunitz soybean trypsin inhibitor

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Abstract

Method of high-sensitivity modulation differential scanning calorimetry was applied for investigation of the kinetically controlled irreversible thermal denaturation of the trypsin inhibitor from soybeans (Kunitz inhibitor, KI) in diluted solution. The measurements were carried out with a temperature-modulation capillary nanocalorimeter designed and produced by the Institute of Biological Instrumentation of the RAS (Pushchino, Russia). An algorithm of the experimental data processing and corresponding software were developed. It was shown that the modulation nanocalorimetry allows one to obtain in one experiment the temperature dependence of the rate constant for irreversible protein denaturation. The temperature dependence of the rate constant and the activation energy of the irreversible denaturation of Kunitz inhibitor were determined. The obtained value of the activation energy (E a = 206 ± 6 kJ mol−1) agrees with independent estimates of this kinetic parameter.

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References

  1. Roberts CJ, Darrington RT, Whitley MB. Irreversible aggregation of recombinant bovine granulocyte-colony stimulating factor (bG-CSF) and implications for predicting protein shelf life. J Pharm Sci. 2003;92:1095–111.

    Article  CAS  Google Scholar 

  2. Roberts CJ. Kinetics of irreversible protein aggregation: analysis of extended Lumry–Eyring models and implications for predicting protein shelf life. J Phys Chem B. 2003;107:1194–207.

    Article  CAS  Google Scholar 

  3. Lumry R, Biltonen R. Thermodynamic and kinetic aspects of protein conformations in relation to the physiological functions. In: Timasheff SN, Fasman GD, editors. Structure and stability of biological macromolecules. Moscow: Mir; 1973. p. 7–173.

    Google Scholar 

  4. Vanosdol WW, Mayorga OL, Freire E. Multifrequency calorimetry of the folding/unfolding transition of cytochrome-C. Biophys J. 1991;59:48–54.

    Article  CAS  Google Scholar 

  5. Reading M. Method and apparatus for modulated differential analysis. Patent USA US5346306. 1994.

  6. Salvetti G, Tombari E, Mikheeva L, Johari GP. The endothermic effects during denaturation of lysozyme by temperature modulated calorimetry and an intermediate reaction equilibrium. J Phys Chem B. 2002;106:6081–7.

    Article  CAS  Google Scholar 

  7. Gmelin E. Classical temperature-modulated calorimetry: a review. Thermochim Acta. 1997;305:1–26.

    Article  Google Scholar 

  8. Imaizumi S, Suzuki K, Hatta I. Ac calorimeter for liquid including suspension of biological-materials. Rev Sci Instrum. 1983;54:1180–5.

    Article  CAS  Google Scholar 

  9. Kraftmakher Y. Modulation calorimetry and related techniques. Phys Rep. 2002;356:1–117.

    Article  CAS  Google Scholar 

  10. Mayorga OL, Rascon AN, Freire E. Multifrequency Calorimetry. Thermochim Acta. 1994;238:309–19.

    Article  CAS  Google Scholar 

  11. Reading M, Elliott D, Hill VL. A new approach to the calorimetric investigation of physical and chemical-transitions. J Therm Anal. 1993;40:949–55.

    Article  CAS  Google Scholar 

  12. Schawe J, Margulies M. Apparatus and method for differential analysis using real and imaginery signal components. European Patent EP0803061. 2004.

  13. Kotelnikov GV, Moiseyeva SP, Mezhburd EV, Krayev VP. Method of separating the sensitive volume of calorimetric cells in a differential titration calorimeter. J Therm Anal Calorim. 2000;62:39–50.

    Article  CAS  Google Scholar 

  14. Kotelnikov GV, Moiseyeva SP, Mezhburd EV, Maevsky EI, Grishina EV. Studying dispersoid systems: method of introducing an injecting needle into calorimetric chamber of capillary titration calorimeter. J Therm Anal Calorim. 2005;81:255–9.

    Article  CAS  Google Scholar 

  15. Kotelnikov GV, Moiseyeva SP, Mezhburd EV. Modulated capillary titration calorimeter: theoretical and experimental studies. J Therm Anal Calorim. 2008;92:631–4.

    Article  CAS  Google Scholar 

  16. Wu YV, Scheraga HA. Studies of soybean trypsin inhibitor. 1. Physicochemical properties. Biochemistry. 1962;1:905–11.

    Article  CAS  Google Scholar 

  17. Kotelnikov GV, Moiseeva SP, Grinberg VY, Khokhlov AR. Modulated capillary differential titration calorimeter. Pribory (Russian). 2009;5:38–44.

    Google Scholar 

  18. Kotelnikov GV, Moiseeva SP. Method of measuring thermal rating on capillary differential volumetric calorimeter. Russian Federation Patent RU2347201. 2009.

  19. Burova TV, Grinberg NV, Lozinskii VI, Moiseeva SP, Kotel’nikov GV, Grinberg VY, Khokhlov AR. Energetics of the binding of Cu(II) ions by thermosensitive copolymers of N-vinylcaprolactam and N-vinylimidazole in different conformational states of macromolecules. Polym Sci Ser A. 2010;52:356–61.

    Article  Google Scholar 

  20. Kalashnikov PA, Kotel’nikov GV, Moiseeva SP, Topchieva IN. Inclusion complexes formed by sodium dodecyl sulfate and cyclodextrin-based nanotubes. Colloid J. 2004;66:542–6.

    Article  CAS  Google Scholar 

  21. Solodovnikov VV. Analysis and synthesis of linear continuous and discrete systems of automatic regulation. Moscow: Mashinostroenie; 1967.

    Google Scholar 

  22. Grinberg VY, Burova TV, Haertle T, Tolstoguzov VB. Interpretation of DSC data on protein denaturation complicated by kinetic and irreversible effects. J Biotechnol. 2000;79:269–80.

    Article  CAS  Google Scholar 

  23. Sanchez-Ruiz JM. Theoretical analysis of Lumry–Eyring models in differential scanning calorimetry. Biophys J. 1992;61:921–35.

    Article  CAS  Google Scholar 

  24. Privalov PL, Potekhin SA. Scanning microcalorimetry in studying temperature-induced changes in proteins. Methods Enzymol. 1986;131:4–51.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This research was supported by the Russian Foundation for Basic Researches (Grant 10-08-00063a).

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Authors and Affiliations

  1. Institute for Biological Instrumentation of the Russian Academy of Sciences, Pushchino, 142290, Moscow Region, Russia

    G. V. Kotelnikov & S. P. Moiseeva

  2. A.N. Nesmeyanov Institute of Organoelement Compounds of the Russian Academy of Sciences, Vavilov Street 28, 119991, Moscow, Russia

    T. V. Burova & N. V. Grinberg

  3. N.M. Emanuel Institute of Biochemical Physics of the Russian Academy of Sciences, Kosygin Street 4, 119991, Moscow, Russia

    A. Ya. Mashkevich, A. S. Dubovik & V. Ya. Grinberg

Authors
  1. G. V. Kotelnikov
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  2. S. P. Moiseeva
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  3. T. V. Burova
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  4. N. V. Grinberg
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  5. A. Ya. Mashkevich
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  6. A. S. Dubovik
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  7. V. Ya. Grinberg
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Correspondence to G. V. Kotelnikov.

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Kotelnikov, G.V., Moiseeva, S.P., Burova, T.V. et al. High-sensitivity modulation differential scanning calorimetry of protein denaturation. J Therm Anal Calorim 114, 531–536 (2013). https://doi.org/10.1007/s10973-013-2966-x

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  • DOI: https://doi.org/10.1007/s10973-013-2966-x

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