Skip to main content
NCBI home page
Biophysical Journal logoLink to Biophysical Journal
. 1995 Dec;69(6):2679–2694. doi: 10.1016/S0006-3495(95)80139-6

Characterizing the secondary hydration shell on hydrated myoglobin, hemoglobin, and lysozyme powders by its vitrification behavior on cooling and its calorimetric glass-->liquid transition and crystallization behavior on reheating.

G Sartor 1, A Hallbrucker 1, E Mayer 1
PMCID: PMC1236505  PMID: 8599674

Abstract

For hydrated metmyoglobin, methemoglobin, and lysozyme powders, the freezable water fraction of between approximately 0.3-0.4 g water/g protein up to approximately 0.7-0.8 g water/g protein has been fully vitrified by cooling at rates up to approximately 1500 K min-1 and the influence of cooling rate characterized by x-ray diffractograms. This vitreous but freezable water fraction started to crystallize at approximately 210 K to cubic ice and at approximately 240 K to hexagonal ice. Measurements by differential scanning calorimetry have shown that this vitreous but freezable water fraction undergoes, on reheating at a rate of 30 K min-1, a glass-->liquid transition with an onset temperature of between approximately 164 and approximately 174 K, with a width of between approximately 9 and approximately 16 degrees and an increase in heat capacity of between approximately 20 and approximately 40 J K-1 (mol of freezable water)-1 but that the glass transition disappears upon crystallization of the freezable water. These calorimetric features are similar to those of water imbibed in the pores of a synthetic hydrogel but very different from those of glassy bulk water. The difference to glassy bulk water's properties is attributed to hydrophilic interaction and H-bonding of the macromolecules' segments with the freezable water fraction, which thereby becomes dynamically modified. Abrupt increase in minimal or critical cooling rate necessary for complete vitrification is observed at approximately 0.7-0.8 g water/g protein, which is attributed to an abrupt increase of water's mobility, and it is remarkably close to the threshold value of water's mobility on a hydrated protein reported by Kimmich et al. (1990, Biophys. J. 58:1183). The hydration level of approximately 0.7-0.8 g water/g protein is approximately that necessary for completing the secondary hydration shell.

Full text

PDF
2679

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bald W. B. On crystal size and cooling rate. J Microsc. 1986 Jul;143(Pt 1):89–102. doi: 10.1111/j.1365-2818.1986.tb02767.x. [DOI] [PubMed] [Google Scholar]
  2. Cooke R., Kuntz I. D. The properties of water in biological systems. Annu Rev Biophys Bioeng. 1974;3(0):95–126. doi: 10.1146/annurev.bb.03.060174.000523. [DOI] [PubMed] [Google Scholar]
  3. Doster W., Bachleitner A., Dunau R., Hiebl M., Lüscher E. Thermal properties of water in myoglobin crystals and solutions at subzero temperatures. Biophys J. 1986 Aug;50(2):213–219. doi: 10.1016/S0006-3495(86)83455-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Doster W., Cusack S., Petry W. Dynamical transition of myoglobin revealed by inelastic neutron scattering. Nature. 1989 Feb 23;337(6209):754–756. doi: 10.1038/337754a0. [DOI] [PubMed] [Google Scholar]
  5. Doster W, Cusack S, Petry W. Dynamic instability of liquidlike motions in a globular protein observed by inelastic neutron scattering. Phys Rev Lett. 1990 Aug 20;65(8):1080–1083. doi: 10.1103/PhysRevLett.65.1080. [DOI] [PubMed] [Google Scholar]
  6. Frauenfelder H., Gratton E. Protein dynamics and hydration. Methods Enzymol. 1986;127:207–216. doi: 10.1016/0076-6879(86)27017-2. [DOI] [PubMed] [Google Scholar]
  7. Goldanskii V. I., Krupyanskii Y. F. Protein and protein-bound water dynamics studied by Rayleigh scattering of Mössbauer radiation (RSMR). Q Rev Biophys. 1989 Feb;22(1):39–92. doi: 10.1017/s003358350000336x. [DOI] [PubMed] [Google Scholar]
  8. Kimmich R., Gneiting T., Kotitschke K., Schnur G. Fluctuations, exchange processes, and water diffusion in aqueous protein systems: A study of bovine serum albumin by diverse NMR techniques. Biophys J. 1990 Nov;58(5):1183–1197. doi: 10.1016/S0006-3495(90)82459-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kuntz I. D., Jr, Kauzmann W. Hydration of proteins and polypeptides. Adv Protein Chem. 1974;28:239–345. doi: 10.1016/s0065-3233(08)60232-6. [DOI] [PubMed] [Google Scholar]
  10. Lounnas V., Pettitt B. M., Phillips G. N., Jr A global model of the protein-solvent interface. Biophys J. 1994 Mar;66(3 Pt 1):601–614. doi: 10.1016/s0006-3495(94)80835-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mayer E., Astl G. Limits of cryofixation as seen by Fourier transform infrared spectra of metmyoglobin azide and carbonyl hemoglobin in vitrified and freeze-concentrated aqueous solution. Ultramicroscopy. 1992 Sep;45(2):185–197. doi: 10.1016/0304-3991(92)90508-h. [DOI] [PubMed] [Google Scholar]
  12. Mayer E. FTIR spectroscopic study of the dynamics of conformational substates in hydrated carbonyl-myoglobin films via temperature dependence of the CO stretching band parameters. Biophys J. 1994 Aug;67(2):862–873. doi: 10.1016/S0006-3495(94)80547-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Olsen K. W. Thermal denaturation procedures for hemoglobin. Methods Enzymol. 1994;231:514–524. doi: 10.1016/0076-6879(94)31035-1. [DOI] [PubMed] [Google Scholar]
  14. Parak F. Correlation of protein dynamics with water mobility: Mössbauer spectroscopy and microwave absorption methods. Methods Enzymol. 1986;127:196–206. doi: 10.1016/0076-6879(86)27016-0. [DOI] [PubMed] [Google Scholar]
  15. Pethig R. Protein-water interactions determined by dielectric methods. Annu Rev Phys Chem. 1992;43:177–205. doi: 10.1146/annurev.pc.43.100192.001141. [DOI] [PubMed] [Google Scholar]
  16. Plattner H., Bachmann L. Cryofixation: a tool in biological ultrastructural research. Int Rev Cytol. 1982;79:237–304. doi: 10.1016/s0074-7696(08)61676-9. [DOI] [PubMed] [Google Scholar]
  17. Poole P. L., Finney J. L. Solid-phase protein hydration studies. Methods Enzymol. 1986;127:284–293. doi: 10.1016/0076-6879(86)27023-8. [DOI] [PubMed] [Google Scholar]
  18. Rothgeb T. M., Gurd F. R. Physical methods for the study of myoglobin. Methods Enzymol. 1978;52:473–486. doi: 10.1016/s0076-6879(78)52052-1. [DOI] [PubMed] [Google Scholar]
  19. Rupley J. A., Careri G. Protein hydration and function. Adv Protein Chem. 1991;41:37–172. doi: 10.1016/s0065-3233(08)60197-7. [DOI] [PubMed] [Google Scholar]
  20. Sartor G., Mayer E. Calorimetric study of crystal growth of ice in hydrated methemoglobin and of redistribution of the water clusters formed on melting the ice. Biophys J. 1994 Oct;67(4):1724–1732. doi: 10.1016/S0006-3495(94)80646-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Sartor G., Mayer E., Johari G. P. Calorimetric studies of the kinetic unfreezing of molecular motions in hydrated lysozyme, hemoglobin, and myoglobin. Biophys J. 1994 Jan;66(1):249–258. doi: 10.1016/S0006-3495(94)80774-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Smith J., Kuczera K., Karplus M. Dynamics of myoglobin: comparison of simulation results with neutron scattering spectra. Proc Natl Acad Sci U S A. 1990 Feb;87(4):1601–1605. doi: 10.1073/pnas.87.4.1601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Srajer V., Reinisch L., Champion P. M. Investigation of laser-induced long-lived states of photolyzed MbCO. Biochemistry. 1991 May 21;30(20):4886–4895. doi: 10.1021/bi00234a008. [DOI] [PubMed] [Google Scholar]
  24. Teeter M. M. Order and disorder in water structure of crystalline proteins. Dev Biol Stand. 1992;74:63–72. [PubMed] [Google Scholar]
  25. Waterman M. R. Spectral characterization of human hemoglobin and its derivatives. Methods Enzymol. 1978;52:456–463. doi: 10.1016/s0076-6879(78)52050-8. [DOI] [PubMed] [Google Scholar]
  26. Yang P. H., Rupley J. A. Protein--water interactions. Heat capacity of the lysozyme--water system. Biochemistry. 1979 Jun 12;18(12):2654–2661. doi: 10.1021/bi00579a035. [DOI] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES