Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 31584
Temperature-dependent Structural Perturbation of Tuna Myoglobin

Authors: Yoshihiro Ochiai


To unveil the mechanism of fast autooxidation of fish myoglobins, the effect of temperature on the structural change of tuna myoglobin was investigated. Purified myoglobin was subjected to preincubation at 5, 20, 50 and 40oC. Overall helical structural decay through thermal treatment up to 95oC was monitored by circular dichroism spectrometry, while the structural changes around the heme pocket was measured by ultraviolet/visible absorption spectrophotometry. As a result, no essential structural change of myoglobin was observed under 30oC, roughly equivalent to their body temperature, but the structure was clearly damaged at 40oC. The Soret band absorption hardly differed irrespective of preincubation temperature, suggesting that the structure around the heme pocket was not perturbed even after thermal treatment.

Keywords: denaturation, myoglobin, stability, tuna.

Digital Object Identifier (DOI):

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1780


[1] S. E. V. Phillips and B.P. Schoenborn, "Neutron diffraction reveals oxygen-histidine hydrogen bond in oxymyoglobin," Nature, vol. 292, pp.81-82, 1981.
[2] J. Vojtechovsky, K. Chu, J. Berendzen, R. M. Sweet, and I. Schlichting, "Crystal structures of myoglobin-ligand complexes at near-atomic resolution," Biophysical Journal, vol. 77, pp.2153-2174, 1999.
[3] G. I. Birnbaum, S. V. Evans, M. Przybylska, and D. R. Rose, "1.70 Å resolution structure of myoglobin from yellowfin tuna. An example of a myoglobin lacking the D helix," Acta Crystallographica, vol. D50, pp.283-289, 1994.
[4] F. Matsuura and K. Hashimoto, "Chemical studies on the red muscle ("chiai") of fishes-X. A new method for determination of myoglobin," Nippon Suisan Gakkaishi, vol. 24, pp.809-815, 1959.
[5] N. Ueki and Y. Ochiai, "Primary structure and thermostability of bigeye tuna myoglobin in relation to those from other scombridae fish," Fisheries Science, vol. 70, pp.875-884, 2004.
[6] N. Ueki, C. J. Chow, and Y. Ochiai, "Characterization of bullet tuna myoglobin with reference to thermostability-structure relationship", Journal of Agricultural and Food Chemistry, vol. 53, pp.4968-4975, 2005.
[7] S. J. Smerdon, S. Krzywda, A. J. Wilkinson, R. E. Brantley, Jr., T. E. Carver, M. S. Hargrove, and J. S. Olsen, "Serine92 (F7) contributes to the control of heme reactivity and stability in myoglobin," Biochemistry, vol. 32, pp.5132-5138, 1993.
[8] Y. Luo, M. S. Kay, and R. L. Baldwin, "Cooperativity of folding of the apomyoglobin pH 4 intermediate studies by glycine and proline mutations," Nature Structure Biology, vol.4, pp.925-930, 1997.
[9] N. Ueki and Y. Ochiai, "Effect of amino acid replacements on the structural stability of fish myoglobin," Journal of Biochemistry, vol.140, pp.649-656, 2006.
[10] Y. Ochiai, N. Ueki and S. Watabe, "Effects of point mutations on the structural stability of tuna myoglobins," Comparative Biochemistry and Physiology, vol. 153B, pp.223-228, 2009.
[11] P. W. Madden, M. J. Babcock, M. E. Vayda, and R. E. Cashon, "Structural and kinetic characterization of myoglobins from eurythermal and stenothermal fish species," Comparative Biochemistry and Physiology, vol.137B, pp.341-350, 2004.
[12] Y. Kitahara, A. Matsuoka, N. Kobayashi, and K. Shikama, "Autooxidation of myoglobin from bigeye tuna fish (Thunnus obesus)," Biochimica Biophysica Acta, vol. 1038, pp.23-28, 1990.
[13] S. Chanthai, M. Ogawa, T. Tamiya, and T. Tsuchiya, "Studies on thermal denaturation of fish myoglobins using differential scanning calorimetry, circular dichroism, and tryptophan fluorescence," Fisheries Science, vol.62, pp.927-932, 1996.
[14] C. J. Chow, Y. Ochiai, S. Watabe, and K. Hashimoto, "Reduced stability and accelerated autooxidation of tuna myoglobin in association with freezing and thawing," Journal of Agricultural and Food Chemistry, vol. 37, pp.1391-1395, 1989.
[15] C. J. Chow, J. C. Wu, P. F. Lee, and Y. Ochiai, "Effects of acid and alkaline pretreatment on the discoloration rates of dark muscle and myoglobin extract of skinned tilapia fillet during iced storage," Fisheries Science vol.75, pp.1481-1488, 2009.
[16] C. J. Chow, "Relationship between the stability and autoxidation of myoglobin," Journal of Agricultural and Food Chemistry, vol.39, pp.22-26, 1991.
[17] Y. Ochiai, Y. Watanabe, N. Uchida, H. Ozawa, S. Ikegami, and S. Watabe, "Thermal denaturation profiles of tuna myoglobin," Bioscience, Biotechnology and Biochemistry, vol.74, pp.1673-1679, 2010.
[18] H. Ozawa, S. Watabe, and Y. Ochiai, "Thermostability of striated and smooth adductor muscle tropomyosins from Yesso scallop Mizuhopecten yessoensis," Journal of Biochemistry, vol.147, pp.823-832, 2010.
[19] B. A. Block, H. Dewar, S. B. Blackwell, T. D. Williams, E. D. Prince, C. J. Farwell, A. Boustany, S. L. H. Teo, A. Seitz, A. Walli, and D. Fudge, "Migratory movements, depth preferences, and thermal biology of Atlantic bluefin tuna," Science, vol.293, pp.1310-1314, 2001.
[20] E. Bismuto, E. Gratton, and D. C. Lamb, "Dynamics of ANS Binding to tuna apomyoglobin measured with fluorescence correlation spectroscopy," Biophysical Journal, vol.81, pp.3510-3521, 2001.
[21] M. Fandrich, V. Forge, K. Buder, M. Kittler, C. M. Dobson, and S. Diekmann, "Myoglobin forms amyloid fibrils by association of unfolded polypeptide segments," Proceedings of the National Academy of Sciences U.S.A., vol.100, pp.15463-15468, 2003.
[22] G. G. Tartaglia, A. P. Pawar, S. Campioni, C. M. Dobson, F. Chiti, and M. Vendruscolo, "Prediction of aggregation-prone regions in structured proteins," Journal of Molecular Biology, vol.380, pp.425-436, 2008.
[23] S. N. Loh, M. S. Kay, and R. L. Baldwin, "Structure and stability of a second molten globule intermediate in the apomyoglobin folding pathway," Proceedings of the National Academy of Sciences U.S.A., vol.92, 5446-5450, 1995.
[24] D. Barrick and R. L. Baldwin, "The molten globule intermediate of apomyoglobin and the process of protein folding," Protein Science, vol.2, pp.869-876, 1993.
[25] E. Bismuto, G. Irace, L. Servillo, A. Giovane, and G. Colonna, "Conformational stability and basal metabolic rate: reexamination of the case of myoglobin," Experientia, vol.40, pp.1400-1401, 1984.
[26] R. E. Cashon, M. E. Vayda, and B. D. Sidell, "Kinetic characterization of myoglobins from vertebrates with vastly different body temperatures," Comparative Biochemistry and Physiology, vol. 117B, 613-620, 1997.
[27] D. J. Marcinek, J. Bonaventura, J. B. Wittenberg, and B. A. Block, "Oxygen affinity and amino acid sequence of myoglobins from endothermic and ectothermic fish," American Journal of Physiology- Regulatory, Integrative and Comparative Physiology, vol.280, pp.R1123-1133, 2001.
[28] S. Hirota, K. Azuma, M. Fukuba, S. Kuroiwa, and N. Funasaki, "Heme reduction by intramolecular electron transfer in cysteine mutant myoglobin under carbon monoxide atmosphere," Biochemistry, vol.44, pp.10322-10327, 2005.
[29] D. Bourgeois, B. Vallone, A. Arcovito, G. Sciara, F. Schotte, P. A. Anfinrud, and M. Brunori, "Extended subnanosecond structural dynamics of myoglobin revealed by Laue crystallography," Proceedings of the National Academy of Sciences U.S.A., vol.103, pp.4924-4929, 2006.
[30] P. Picotti, A. Marabotti, A. Negro, V. Musi, B. Spolaore, M. Zambonin, and A. Fontana, "Modulation of structural integrity of helix F in apomyoglobin by single amino acid replacements," Protein Science, vol.13, pp.1572-1585, 2004.
[31] A. E. Dyuysekina, D. A. Dolgikh, E. N. Samatova Baryshnikova, E. I. Tiktopulo, V. A. Balobanov, and V. E. Bychkova, "pH-induced equilibrium unfolding of apomyoglobin: substitutions at conserved Trp14 and Met131 and non-conserved Val17 positions," Biochemistry (Moscow), vol.73, pp.693-701, 2008.
[32] R. Musto, M. G. Bigotti, C. Travaglini-Allocatelli, M. Brunori, and F. Cutruzzola, "Folding of Aplysia limacina apomyoglobin involves an intermediate in common with other evolutionarily distant globins," Biochemistry, vol.43, pp.230-236, 2004.
[33] M. S. Hargrove, E. W. Singleton, M. L. Quillin, L. A. Ortiz, G. N. Phillips Jr., J. S. Olsen, and A. J. Mathews, "His64(E7)  Tyr apomyoglobin as a reagent for measuring rates of hemin dissociation," Journal of Biological Chemistry, vol.269, pp.4207-4214, 1994.
[34] E. Bismuto, E. Di Maggio, S. Pleus, M. Sikor, C. Rocker, G. H. Nienhaus, and D. C. Lamb, "Molecular dynamics simulation of the acidic compact state of apomyoglobin from yellowfin tuna," Proteins, vol.74, pp.273-290, 2008.
[35] M. Dametto and A. E. Cardenas, "Computer simulations of the refolding of sperm whale apomyoglobin from high-temperature denaturated state," Journal of Physical Chemistry B, vol.112, pp.9501-9506, 2008.