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Home / Equine Research Bank / Biophys J / Surface plasmon resonance studies of complex formation betwe...
Biophysical journal1996; 71(2); 858-867; doi: 10.1016/S0006-3495(96)79287-1

Surface plasmon resonance studies of complex formation between cytochrome c and bovine cytochrome c oxidase incorporated into a supported planar lipid bilayer. II. Binding of cytochrome c to oxidase-containing cardiolipin/phosphatidylcholine membranes.

Abstract: Complex formation between horse heart cytochrome c (cyt c) and bovine cytochrome c oxidase (cco) incorporated into a supported planar egg phosphatidylcholine membrane containing varying amounts of cardiolipin (CL) (0-20 mol%) has been studied under low (10 mM) and medium (160 mM) ionic strength conditions by surface plasmon resonance (SPR) spectroscopy. Both specific and nonspecific modes of cyt c binding are observed. The dissociation constant of the specific interaction between cyt c and cco increases from approximately 6.5 microM at low ionic strength to 18 microM at medium ionic strength, whereas the final saturation level of bound protein is independent of salt concentration and corresponds to approximately 53% of the total cco molecules present in the membrane. This suggests a 1:1 binding stoichiometry between the two proteins. The nonspecific binding component is governed by electrostatic interactions between cyt c and the membrane lipids and results in a partially ionic strength-reversible protein-membrane association. Thus, hydrophobic interactions between cyt c and the membrane, which are the predominant mode of binding in the absence of cco, are greatly suppressed. Both the amount of nonspecifically bound protein and the binding affinity can be varied over a broad range by changing the ionic strength and the extent of CL incorporation into the membrane. Under conditions approximating the physiological state in the mitochondrion (i.e., 20 mol% CL and medium ionic strength), 1-1.5 cyt c molecules are bound to the lipid phase per molecule of cco, with a dissociation constant of 0.1 microM. The possible physiological significance of these observations is discussed.
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Publication Date: 1996-08-01 PubMed ID: 8842224PubMed Central: PMC1233542DOI: 10.1016/S0006-3495(96)79287-1Google Scholar: Lookup
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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.

The research article is about the study of the interaction between horse heart cytochrome c and bovine cytochrome c oxidase under varying conditions, using surface plasmon resonance spectroscopy.

Introduction and Methodology

  • The research focuses on understanding the complex formation between two proteins, horse heart cytochrome c (cyt c) and bovine cytochrome c oxidase (cco).
  • The proteins are incorporated into a supported planar egg phosphatidylcholine membrane with varying cardiolipin (CL) content. This variation ranges from 0-20 mol%.
  • The study is conducted under different ionic strength conditions (10 mM and 160 mM) using surface plasmon resonance (SPR) spectroscopy.

Observations

  • The interaction is observed in two ways – specific and nonspecific.
  • The dissociation constant of the specific interaction between the two proteins increases when the ionic strength increases, whereas the final saturation level of the bound protein is independent of salt concentration.
  • The number of protein-cco complexes corresponds to approximately 53% of the total cco molecules in the membrane, indicating a 1:1 binding ratio between the cyt c and cco proteins.
  • In the case of nonspecific binding, interaction is governed by electrostatic interactions between the cyt c and lipids in the membrane, resulting in a partly reversible protein-membrane connection, which is impacted by the ionic strength.
  • The researchers found that hydrophobic interactions between cyt c and the membrane, which are the primary binding mode when cco is absent, are significantly reduced.

Finding and Conclusion

  • Variations in binding affinity and the amount of nonspecifically bound protein are possible by changing the ionic strength and CL content in the membrane.
  • Under conditions that resemble physiological states in the mitochondrion (20 mol% CL and medium ionic strength), the researchers found that between 1 and 1.5 cyt c molecules are bound to the lipid phase for each cco molecule, with a dissociation constant of 0.1 microM.
  • The physiological implications of these interactions are also discussed, offering insight into the function and behavior of these components within the biological cell environment.

Cite This Article

APA
Salamon Z, Tollin G. (1996). Surface plasmon resonance studies of complex formation between cytochrome c and bovine cytochrome c oxidase incorporated into a supported planar lipid bilayer. II. Binding of cytochrome c to oxidase-containing cardiolipin/phosphatidylcholine membranes. Biophys J, 71(2), 858-867. https://doi.org/10.1016/S0006-3495(96)79287-1

Publication

Biophysical journal
ISSN: 0006-3495
NlmUniqueID: 0370626
Country: United States
Language: English
Volume: 71
Issue: 2
Pages: 858-867

Researcher Affiliations

Salamon, Z
  • Department of Biochemistry, University of Arizona, Tucson 85721, USA.
Tollin, G

    MeSH Terms

    • Animals
    • Cardiolipins
    • Cattle
    • Cytochrome c Group / chemistry
    • Cytochrome c Group / metabolism
    • Electron Transport Complex IV / chemistry
    • Electron Transport Complex IV / metabolism
    • Horses
    • Kinetics
    • Lipid Bilayers
    • Models, Structural
    • Phosphatidylcholines
    • Protein Binding

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    Citations

    This article has been cited 11 times.
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