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Home / Equine Research Bank / Protein Sci / Volume changes of the molten globule transitions of horse he...
Protein science : a publication of the Protein Society1995; 4(7); 1426-1429; doi: 10.1002/pro.5560040717

Volume changes of the molten globule transitions of horse heart ferricytochrome c: a thermodynamic cycle.

Abstract: Volume changes among the unfolded (U), native (N), and molten globule (MG) conformations of horse heart ferricytochrome c have been measured. U to N (pH 2 to pH 7) was determined in the absence of added salt to be -136 +/- 5 mL/mol protein. U to MG (pH 2, no added salt to pH 2, 0.5 M KCl) yielded + 100 +/- 6 mL/mol. MG to N was broken into two steps, N to NClx at pH 7 by addition of buffered KCl to buffered protein lacking added salt (NClx = N interacting with an unknown number, X, of chloride ions), and MG to NClx by jumping MG at pH 2 in 0.5 M KCl to pH7 at the same salt concentration. The delta V of N to NClx was -30.9 +/- 1.4 mL/mol protein, whereas MG to NClx entailed a delta V of -235 +/- 6 mL/mol. Within experimental error, the results add up to zero for a complete thermodynamic cycle. We believe this to be the first volumetric cycle to have been measured for the conformational transitions of a protein. The results are discussed in terms of hydration contributions from deprotonation of the protein, other hydration effects, and the formation and/or enlargement of packing defects in the protein's tertiary structure during the steps of folding.
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Publication Date: 1995-07-01 PubMed ID: 7670384PubMed Central: PMC2143161DOI: 10.1002/pro.5560040717Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research investigates and measures the volume changes that occur during the different conformational transitions (changes in shape) of the ferricytochrome c protein present in horse hearts.

Study Overview

  • The paper presents an examination of the volume changes among three state changes of horse heart ferricytochrome c. These states are the Unfolded (U), Native (N), and molten globule (MG) conformations
  • If ferricytochrome c transitions from U to N or U to MG, and eventually N to NClx then MG to NClx, the collective volumetric changes amount to zero, creating a thermodynamic cycle. This is exceptional as this is possibly the first-ever volumetric cycle measured for protein conformational transitions.

Research Methodology and Results

  • Transition of U to N (i.e., transition from pH 2 to pH 7 without any additional salt) measured a volume change of -136 +/- 5 mL/mol protein.
  • Transition of U to MG (i.e., movement from pH 2 with no additional salt to pH 2, with 0.5 M KCl) resulted in a volume increase of +100 +/- 6 mL/mol.
  • The researchers broke the transition from MG to N into two points: first, N to NClx at pH 7 by adding buffered KCl to buffered protein lacking added salt (where NClx = N interacting with an unknown number, X, of chloride ions), and second, MG to NClx by jumping MG at pH 2 in 0.5 M KCl to pH7 at the same salt concentration.
  • The volume change (delta V) of N to NClx was -30.9 +/- 1.4 mL/mol protein, whereas MG to NClx entailed a delta V of -235 +/- 6 mL/mol.

Impact and Conclusion of the Study

  • This research is significant as it allegedly presents the first volumetric cycle measured for the conformational transitions of a protein.
  • The findings could provide insights into the hydration influences from deprotonation of the protein, other hydration impacts, and the development or enlargement of packing defects in the protein’s tertiary structure during different folding stages.
  • Understanding these volume changes and the conditions that bring them about is vital for understanding the specific transitions, how proteins might react under different conditions, and could have implications for protein engineering and biopharmaceutics.

Cite This Article

APA
Foygel K, Spector S, Chatterjee S, Kahn PC. (1995). Volume changes of the molten globule transitions of horse heart ferricytochrome c: a thermodynamic cycle. Protein Sci, 4(7), 1426-1429. https://doi.org/10.1002/pro.5560040717

Publication

Protein science : a publication of the Protein Society
ISSN: 0961-8368
NlmUniqueID: 9211750
Country: United States
Language: English
Volume: 4
Issue: 7
Pages: 1426-1429

Researcher Affiliations

Foygel, K
  • Department of Biochemistry and Microbiology, Rutgers University, New Brunswick, New Jersey 08903, USA.
Spector, S
    Chatterjee, S
      Kahn, P C

        MeSH Terms

        • Animals
        • Chemical Phenomena
        • Chemistry, Physical
        • Cytochrome c Group / chemistry
        • Horses
        • Hydrogen-Ion Concentration
        • Myocardium / chemistry
        • Potassium Chloride / pharmacology
        • Protein Conformation
        • Protein Folding
        • Thermodynamics

        References

        This article includes 29 references
        1. Babul J, Stellwagen E. Participation of the protein ligands in the folding of cytochrome c.. Biochemistry 1972 Mar 28;11(7):1195-200.
          pubmed: 5062485doi: 10.1021/bi00757a013google scholar: lookup
        2. Lee B, Richards FM. The interpretation of protein structures: estimation of static accessibility.. J Mol Biol 1971 Feb 14;55(3):379-400.
          pubmed: 5551392doi: 10.1016/0022-2836(71)90324-xgoogle scholar: lookup
        3. Chothia C. Hydrophobic bonding and accessible surface area in proteins.. Nature 1974 Mar 22;248(446):338-9.
          pubmed: 4819639doi: 10.1038/248338a0google scholar: lookup
        4. Weber G, Tanaka F, Okamoto BY, Drickamer HG. The effect of pressure on the molecular complex of isoalloxazine and adenine.. Proc Natl Acad Sci U S A 1974 Apr;71(4):1264-6.
          pubmed: 4524637doi: 10.1073/pnas.71.4.1264google scholar: lookup
        5. Stellwagen E, Babul J. Stabilization of the globular structure of ferricytochrome c by chloride in acidic solvents.. Biochemistry 1975 Nov 18;14(23):5135-40.
          pubmed: 41doi: 10.1021/bi00694a018google scholar: lookup
        6. Shaw RW, Hartzell CR. Hydrogen ion titration of horse heart ferricytochrome c.. Biochemistry 1976 May 4;15(9):1909-14.
          pubmed: 5119doi: 10.1021/bi00654a018google scholar: lookup
        7. Bernstein FC, Koetzle TF, Williams GJ, Meyer EF Jr, Brice MD, Rodgers JR, Kennard O, Shimanouchi T, Tasumi M. The Protein Data Bank: a computer-based archival file for macromolecular structures.. J Mol Biol 1977 May 25;112(3):535-42.
          pubmed: 875032doi: 10.1016/s0022-2836(77)80200-3google scholar: lookup
        8. Visser AJ, Li TM, Drickamer HG, Weber G. Volume changes in the formation of internal complexes of flavinyltryptophan peptides.. Biochemistry 1977 Nov 1;16(22):4883-6.
          pubmed: 911796doi: 10.1021/bi00641a021google scholar: lookup
        9. Taborsky G, McCollum K. Phosphate binding by cytochrome c. Specific binding site involved in the formation and reactivity of a complex of ferricytochrome c, ferrous ion, and phosphate.. J Biol Chem 1979 Aug 10;254(15):7069-75.
          pubmed: 222755
        10. Osheroff N, Brautigan DL, Margoliash E. Mapping of anion binding sites on cytochrome c by differential chemical modification of lysine residues.. Proc Natl Acad Sci U S A 1980 Aug;77(8):4439-43.
          pubmed: 6254024doi: 10.1073/pnas.77.8.4439google scholar: lookup
        11. Dyson HJ, Beattie JK. Spin state and unfolding equilibria of ferricytochrome c in acidic solutions.. J Biol Chem 1982 Mar 10;257(5):2267-73.
          pubmed: 6277891
        12. Kahn PC, Briehl RW. The absence of volume change in the gelation of hemoglobin-S.. J Biol Chem 1982 Oct 25;257(20):12209-13.
          pubmed: 7118939
        13. Ohgushi M, Wada A. 'Molten-globule state': a compact form of globular proteins with mobile side-chains.. FEBS Lett 1983 Nov 28;164(1):21-4.
          pubmed: 6317443doi: 10.1016/0014-5793(83)80010-6google scholar: lookup
        14. Dolgikh DA, Kolomiets AP, Bolotina IA, Ptitsyn OB. 'Molten-globule' state accumulates in carbonic anhydrase folding.. FEBS Lett 1984 Jan 2;165(1):88-92.
          pubmed: 6420185doi: 10.1016/0014-5793(84)80020-4google scholar: lookup
        15. Eisenberg D, Wilcox W, McLachlan AD. Hydrophobicity and amphiphilicity in protein structure.. J Cell Biochem 1986;31(1):11-7.
          pubmed: 3722276doi: 10.1002/jcb.240310103google scholar: lookup
        16. Trewhella J, Carlson VA, Curtis EH, Heidorn DB. Differences in the solution structures of oxidized and reduced cytochrome c measured by small-angle X-ray scattering.. Biochemistry 1988 Feb 23;27(4):1121-5.
          pubmed: 2835084doi: 10.1021/bi00404a007google scholar: lookup
        17. Bychkova VE, Pain RH, Ptitsyn OB. The 'molten globule' state is involved in the translocation of proteins across membranes?. FEBS Lett 1988 Oct 10;238(2):231-4.
          pubmed: 3049159doi: 10.1016/0014-5793(88)80485-xgoogle scholar: lookup
        18. Liu GY, Grygon CA, Spiro TG. Ionic strength dependence of cytochrome c structure and Trp-59 H/D exchange from ultraviolet resonance Raman spectroscopy.. Biochemistry 1989 Jun 13;28(12):5046-50.
          pubmed: 2548599doi: 10.1021/bi00438a022google scholar: lookup
        19. Kuwajima K. The molten globule state as a clue for understanding the folding and cooperativity of globular-protein structure.. Proteins 1989;6(2):87-103.
          pubmed: 2695928doi: 10.1002/prot.340060202google scholar: lookup
        20. Goto Y, Takahashi N, Fink AL. Mechanism of acid-induced folding of proteins.. Biochemistry 1990 Apr 10;29(14):3480-8.
          pubmed: 2162192doi: 10.1021/bi00466a009google scholar: lookup
        21. Christensen H, Pain RH. Molten globule intermediates and protein folding.. Eur Biophys J 1991;19(5):221-9.
          pubmed: 2060495doi: 10.1007/BF00183530google scholar: lookup
        22. Jeng MF, Englander SW. Stable submolecular folding units in a non-compact form of cytochrome c.. J Mol Biol 1991 Oct 5;221(3):1045-61.
          pubmed: 1658332doi: 10.1016/0022-2836(91)80191-vgoogle scholar: lookup
        23. Goto Y, Nishikiori S. Role of electrostatic repulsion in the acidic molten globule of cytochrome c.. J Mol Biol 1991 Dec 5;222(3):679-86.
          pubmed: 1660930doi: 10.1016/0022-2836(91)90504-ygoogle scholar: lookup
        24. Myer YP, Saturno AF. Horse heart ferricytochrome c: conformation and heme configuration of high ionic strength acidic forms.. J Protein Chem 1991 Oct;10(5):481-94.
          pubmed: 1665977doi: 10.1007/BF01025476google scholar: lookup
        25. Haynie DT, Freire E. Structural energetics of the molten globule state.. Proteins 1993 Jun;16(2):115-40.
          pubmed: 8332604doi: 10.1002/prot.340160202google scholar: lookup
        26. Goto Y, Hagihara Y, Hamada D, Hoshino M, Nishii I. Acid-induced unfolding and refolding transitions of cytochrome c: a three-state mechanism in H2O and D2O.. Biochemistry 1993 Nov 9;32(44):11878-85.
          pubmed: 8218260doi: 10.1021/bi00095a017google scholar: lookup
        27. Ybe JA, Kahn PC. Slow-folding kinetics of ribonuclease-A by volume change and circular dichroism: evidence for two independent reactions.. Protein Sci 1994 Apr;3(4):638-49.
          pubmed: 8003982doi: 10.1002/pro.5560030412google scholar: lookup
        28. Kasarda DD. Dilution volume changes of some purine and pyrimidine compounds.. Biochim Biophys Acta 1970 Oct 15;217(2):535-8.
          pubmed: 5473201doi: 10.1016/0005-2787(70)90552-6google scholar: lookup
        29. Richards FM. The interpretation of protein structures: total volume, group volume distributions and packing density.. J Mol Biol 1974 Jan 5;82(1):1-14.
          pubmed: 4818482doi: 10.1016/0022-2836(74)90570-1google scholar: lookup

        Citations

        This article has been cited 7 times.
        1. Kahn PC. The measurement of volume change by capillary dilatometry. Protein Sci 2019 Jun;28(6):1135-1142.
          doi: 10.1002/pro.3626pubmed: 30993790google scholar: lookup
        2. El Kadi N, Taulier N, Le Huérou JY, Gindre M, Urbach W, Nwigwe I, Kahn PC, Waks M. Unfolding and refolding of bovine serum albumin at acid pH: ultrasound and structural studies. Biophys J 2006 Nov 1;91(9):3397-404.
          doi: 10.1529/biophysj.106.088963pubmed: 16861279google scholar: lookup
        3. Watanabe M, Kobashigawa Y, Aizawa T, Demura M, Nitta K. A non-native alpha-helix is formed in the beta-sheet region of the molten globule state of canine milk lysozyme. Protein J 2004 Jul;23(5):335-42.
          doi: 10.1023/b:jopc.0000032653.30096.41pubmed: 15328889google scholar: lookup
        4. DeVane R, Ridley C, Larsen RW, Space B, Moore PB, Chan SI. A molecular dynamics method for calculating molecular volume changes appropriate for biomolecular simulation. Biophys J 2003 Nov;85(5):2801-7.
          doi: 10.1016/S0006-3495(03)74703-1pubmed: 14581185google scholar: lookup
        5. Valdez D, Le Huérou JY, Gindre M, Urbach W, Waks M. Hydration and protein folding in water and in reverse micelles: compressibility and volume changes. Biophys J 2001 Jun;80(6):2751-60.
          doi: 10.1016/S0006-3495(01)76243-1pubmed: 11371450google scholar: lookup
        6. Kobashigawa Y, Sakurai M, Nitta K. Effect of hydrostatic pressure on unfolding of alpha-lactalbumin: volumetric equivalence of the molten globule and unfolded state. Protein Sci 1999 Dec;8(12):2765-72.
          doi: 10.1110/ps.8.12.2765pubmed: 10631994google scholar: lookup
        7. Kornblatt JA, Kornblatt MJ, Rajotte I, Hoa GH, Kahn PC. Thermodynamic volume cycles for electron transfer in the cytochrome c oxidase and for the binding of cytochrome c to cytochrome c oxidase. Biophys J 1998 Jul;75(1):435-44.
          doi: 10.1016/S0006-3495(98)77531-9pubmed: 9649404google scholar: lookup
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