Slab Gel Electrophoresis Performed at High Hydrostatic Pressure: A New Approach for the Study of Oligomeric Proteins

  • Alejandro A. Paladini
  • Jerson L. Silva
  • Gregorio Weber
Conference paper


In the last few years several studies have demonstrated the reversible dissociation of oligomeric proteins under pressure (for review see: Heremans, 1982; Weber and Drickamer, 1983). The dissociating effects of pressure are generally ascribed to the intersubunit surfaces and the restrictive effect of the covalent bond architecture of the protein. The pressure dissociation has been detected either by indirect or direct methods. Indirect methods, such as hybridization studies (Jaenicke and Koberstein, 1971) and activity measurements (Penniston, 1971; Schade et al., 1980; Seifert et al., 1985) have been employed with some success. These indirect methods are valuable in revealing qualitatively the involvement of protein dissociation in the observed effect of pressure. However, the stoichiometry of dissociation and the thermodynamic parameters (volume change and dissociation constant) cannot be unambiguously determined. Methods which directly reveal the state of association are more suitable for a thermodynamic approach to the pressure dissociation, although they are not immune to experimental problems.


High Hydrostatic Pressure Oligomeric Protein Pressure Dissociation Pressure Bomb Single Chain Protein 
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  1. Engelborghs, Y., Heremans, K. A., DeMaeyer, L., and Hoebeke, J., 1976, Effect of Temperature and Pressure on Polymerization Equilibrium of Neuronal Microtubules, Nature, 256:686.CrossRefGoogle Scholar
  2. Heremans, K. A., 1982, High Pressure Effects on Proteins and Other Biomolecules, Ann. Rev. Biophys. and Bioeng., 11:1.CrossRefGoogle Scholar
  3. Hawley, S. A., and Mitchell, R. M., 1975, An Electrophoretic Study of Reversible Protein Denaturation: Chymotrypsinogen at High Pressures, Biochemistry, 14:3257.PubMedCrossRefGoogle Scholar
  4. Hogberg-Raibaud, A., and Goldberg, M. E., 1977, Isolation and Characterization of Independently Folding Regions of the β Chain of Escherichia coli Tryptophan Synthetase, Biochemistry, 16:4014.PubMedCrossRefGoogle Scholar
  5. Jaenicke, R., and Koberstein, R., 1971, High Pressure Dissociation of Lactic Dehydrogenase, FEBS Lett., 17:351.PubMedCrossRefGoogle Scholar
  6. Josephs, R., and Harrington, W. F., 1967, An Unusual Pressure Dependence for a Reversibly Associating Protein System; Sedimentation Studies on Myosin, Proc. Natl. Acad. Sci. U.S.A., 58:1587.PubMedCrossRefGoogle Scholar
  7. King, L., and Weber, G., 1986, Conformational Drift of Dissociated Lactate Dehydrogenases, Biochemistry, 25:3632.PubMedCrossRefGoogle Scholar
  8. Miles, E. W., and Moriguchi, 1977, Tryptophan Synthase of Escherichia coli Removal of Pyridoxal 5′-Phosphate and Separation of the α and β2 Subunits, J. Biol. Chem., 252:6594.PubMedGoogle Scholar
  9. Neuman, R. C., Kauzmann, W., and Zipp, A., 1973, Pressure Dependence of Weak Acid Ionization in Aqueous Buffers, J. Phys. Chem., 77:2687.CrossRefGoogle Scholar
  10. Paladini, A.A., and Weber, G., 1981a, Pressure-Induced Reversible Dissociation of Enolase, Biochemistry, 20:2587.PubMedCrossRefGoogle Scholar
  11. Paladini, A. A., and Weber, G., 1981b, Absolute Measurements of Fluorescence Polarization at High Pressures, Rev. Sci. Instrum., 52:419.CrossRefGoogle Scholar
  12. Payens, T.A.J., and Heremans, K.A.H., 1969, Effect of Pressure on the Temperature-Dependent Association of B-Casein, Biopolymers, 8:335.PubMedCrossRefGoogle Scholar
  13. Penniston, J. T., 1971, High Hydrostatic Pressure and Enzymic Activity: Inhibition of Multimeric Enzymes by Dissociation, Arch. Biochem. Biophys., 142:322.PubMedCrossRefGoogle Scholar
  14. Royer, C. A., Weber, G., Daly, T. J., and Matthews, K. S., 1986, Dissociation of the Lactose Repressor Protein Tetramer Using High Pressure, Biochemistry, 25:8308.PubMedCrossRefGoogle Scholar
  15. Schade, B. C., Rudolph, R., Ludemann, H. D., and Jaenicke, R., 1980, Reversible High-Pressure Dissociation of Lactic Dehydrogenase from Pig, Biochemistry, 19:1121.PubMedCrossRefGoogle Scholar
  16. Seifert, T., Bartholmes, P., and Jaenicke, R., 1985, Influence of Cofactor Pyridoxal 5′-Phosphate on Reversible High-Pressure Denaturation of Isolated β2 Dimer of Tryptophan Synthase Bienzyme Complex from Escherichia coli, Biochemistry, 24:339.PubMedCrossRefGoogle Scholar
  17. Silva, J. L., Miles, E. W., and Weber, G., 1986, Pressure Dissociation and Conformational Drift of the β Dimer of Tryptophan Synthase, Biochemistry, 25:5780.PubMedCrossRefGoogle Scholar
  18. Verjovski-Almeida, S., Kurtenbach, E., Amorim, A. F., and Weber, G., 1986, Pressure-Induced Dissociation of Solubilized Sarcoplasmic Reticulum ATPase, J. Biol. Chem., 261:9872.PubMedGoogle Scholar
  19. Weber, G., and Drickamer, H. G., 1983, The Effect of High Pressure Upon Proteins and other Biomolecules, Quarterly Review of Biophysics, 16:89.CrossRefGoogle Scholar
  20. Weber, K., and Osborn, M., 1969, The Reliability of Molecular Weight Determinations by Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis, J. Biol. Chem., 244:4406.PubMedGoogle Scholar
  21. Zipp, A., and Kauzmann, W., 1973, Pressure Denaturation of Metmyoglobin, Biochemistry, 12:4217.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1989

Authors and Affiliations

  • Alejandro A. Paladini
    • 1
  • Jerson L. Silva
    • 2
  • Gregorio Weber
    • 2
  1. 1.Ingebi-Conicet, Facultad de Ciencias Exactas y NaturalesUniversidad de Buenos AiresArgentina
  2. 2.Department of Biochemistry, School of Chemical SciencesUniversity of IllinoisUrbanaUSA

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