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Biochem J. 1989 Sep 1; 262(2): 591–596.
PMCID: PMC1133309

Ionic-strength-dependence of the oxidation of native and pyridoxal 5'-phosphate-modified cytochromes c by cytochrome c oxidase.


The ionic-strength-dependences of the rate constants (log k plotted versus square root of 1) for oxidation of native and pyridoxal 5'-phosphate-modified cytochromes c by three different preparations of cytochrome c oxidase have complex non-linear character, which may be explained on the basis of present knowledge of the structure of the oxidase and the monomer-dimer equilibrium of the enzyme. The wave-type curve (with a minimum and a maximum) for oxidation of native cytochrome c by purified cytochrome c oxidase depleted of phospholipids may reflect consecutively inhibition of oxidase monomers (initial descending part), competition between this inhibition and dimer formation, resulting in increased activity (second part with positive slope), and finally inhibition of oxidase dimers (last descending part of the curve). The dependence of oxidation of native cytochrome c by cytochrome c oxidase reconstituted into phospholipid vesicles is a curve with a maximum, without the initial descending part described above. This may reflect the lack of pure monomers in the vesicles, where equilibrium is shifted to dimers even at low ionic strength. Subunit-III-depleted cytochrome c oxidase does not exhibit the maximum seen with the other two enzyme preparations. This may mean that removal of subunit III hinders dimer formation. The charge interactions of each of the cytochromes c (native or modified) with the three cytochrome c oxidase preparations are similar, as judged by the similar slopes of the linear dependences at I values above the optimal one. This shows that subunit III and the phospholipid membrane do not seem to be involved in the specific charge interaction of cytochrome c oxidase with cytochrome c.

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  • Koppenol WH, Vroonland CA, Braams R. The electric potential field around cytochrome c and the effect of ionic strength on reaction rates of horse cytochrome c. Biochim Biophys Acta. 1978 Sep 7;503(3):499–508. [PubMed]
  • Koppenol WH. Effect of a molecular dipole on the ionic strength dependence of a biomolecular rate constant. Identification of the site of reaction. Biophys J. 1980 Mar;29(3):493–507. [PMC free article] [PubMed]
  • Koppenol WH, Margoliash E. The asymmetric distribution of charges on the surface of horse cytochrome c. Functional implications. J Biol Chem. 1982 Apr 25;257(8):4426–4437. [PubMed]
  • Staudenmayer N, Smith MB, Smith HT, Spies FK, Jr, Millett F. An enzyme kinetics and 19F nuclear magnetic resonance study of selectively trifluoroacetylated cytochrome c derivatives. Biochemistry. 1976 Jul 27;15(15):3198–3205. [PubMed]
  • Staudenmayer N, Ng S, Smith MB, Millett F. Effect of specific trifluoroacetylation of individual cytochrome c lysines on the reaction with cytochrome oxidase. Biochemistry. 1977 Feb 22;16(4):600–604. [PubMed]
  • Smith HT, Staudenmayer N, Millett F. Use of specific lysine modifications to locate the reaction site of cytochrome c with cytochrome oxidase. Biochemistry. 1977 Nov 15;16(23):4971–4974. [PubMed]
  • Ferguson-Miller S, Brautigan DL, Margoliash E. Definition of cytochrome c binding domains by chemical modification. III. Kinetics of reaction of carboxydinitrophenyl cytochromes c with cytochrome c oxidase. J Biol Chem. 1978 Jan 10;253(1):149–159. [PubMed]
  • Osheroff N, Brautigan DL, Margoliash E. Definition of enzymic interaction domains on cytochrome c. Purification and activity of singly substituted carboxydinitrophenyl-lysine 7, 25, 73, 86, and 99 cytochromes c. J Biol Chem. 1980 Sep 10;255(17):8245–8251. [PubMed]
  • Rieder R, Bosshard HR. The cytochrome c oxidase binding site on cytochrome c. Differential chemical modification of lysine residues in free and oxidase-bound cytochrome c. J Biol Chem. 1978 Sep 10;253(17):6045–6053. [PubMed]
  • Millett F, Darley-Usmar V, Capaldi RA. Cytochrome c is cross-linked to subunit II of cytochrome c oxidase by a water-soluble carbodiimide. Biochemistry. 1982 Aug 3;21(16):3857–3862. [PubMed]
  • Millett F, de Jong C, Paulson L, Capaldi RA. Identification of specific carboxylate groups on cytochrome c oxidase that are involved in binding cytochrome c. Biochemistry. 1983 Feb 1;22(3):546–552. [PubMed]
  • Capaldi RA, Malatesta F, Darley-Usmar VM. Structure of cytochrome c oxidase. Biochim Biophys Acta. 1983 Jul 15;726(2):135–148. [PubMed]
  • Brunori M, Antonini G, Malatesta F, Sarti P, Wilson MT. Cytochrome-c oxidase. Subunit structure and proton pumping. Eur J Biochem. 1987 Nov 16;169(1):1–8. [PubMed]
  • Nałeçz KA, Bolli R, Ludwig B, Azzi A. The role of subunit III in bovine cytochrome c oxidase. Comparison between native, subunit III-depleted and Paracoccus denitrificans enzymes. Biochim Biophys Acta. 1985 Jul 17;808(2):259–272. [PubMed]
  • Veerman EC, Wilms J, Casteleijn G, Van Gelder BF. The pre-steady state reaction of ferrocytochrome c with the cytochrome c-cytochrome aa3 complex. Biochim Biophys Acta. 1980 Mar 7;590(1):117–127. [PubMed]
  • Wilms J, Dekker HL, Boelens R, van Gelder BF. The effect of pH and ionic strength on the pre-steady-state reaction of cytochrome c and cytochrome aa3. Biochim Biophys Acta. 1981 Aug 12;637(1):168–176. [PubMed]
  • Veerman EC, Wilms J, Dekker HL, Muijsers AO, van Buuren KJ, van Gelder BF, Osheroff N, Speck SH, Margoliash E. The presteady state reaction of chemically modified cytochromes c with cytochrome oxidase. J Biol Chem. 1983 May 10;258(9):5739–5745. [PubMed]
  • Sinjorgo KM, Steinebach OM, Dekker HL, Muijsers AO. The effects of pH and ionic strength on cytochrome c oxidase steady-state kinetics reveal a catalytic and a non-catalytic interaction domain for cytochrome c. Biochim Biophys Acta. 1986 Jun 10;850(1):108–115. [PubMed]
  • Bolgiano B, Smith L, Davies HC. Kinetics of the interaction of the cytochrome c oxidase of Paracoccus denitrificans with its own and bovine cytochrome c. Biochim Biophys Acta. 1988 Apr 22;933(2):341–350. [PubMed]
  • Yu C, Yu L, King TE. Studies on cytochrome oxidase. Interactions of the cytochrome oxidase protein with phospholipids and cytochrome c. J Biol Chem. 1975 Feb 25;250(4):1383–1392. [PubMed]
  • Casey RP, Chappell JB, Azzi A. Limited-turnover studies on proton translocation in reconstituted cytochrome c oxidase-containing vesicles. Biochem J. 1979 Jul 15;182(1):149–156. [PMC free article] [PubMed]
  • Bill K, Azzi A. An active cytochrome c oxidase depleted of subunit III prepared by covalent chromatography on yeast cytochrome c. Biochem Biophys Res Commun. 1982 Jun 30;106(4):1203–1209. [PubMed]
  • Atanasov BP, Mitovska MI, Mancheva IN, Kossekova GP, Tchorbanov BP, Christova P, Dancheva KI. Pyridoxal phosphate modified cytochromes c. Identification and electron transfer properties. Biochim Biophys Acta. 1984 Jun 26;765(3):329–339. [PubMed]
  • Nałecz KA, Bolli R, Azzi A. Preparation of monomeric cytochrome C oxidase: its kinetics differ from those of the dimeric enzyme. Biochem Biophys Res Commun. 1983 Jul 29;114(2):822–828. [PubMed]
  • Bolli R, Nałecz KA, Azzi A. The aggregation state of bovine heart cytochrome c oxidase and its kinetics in monomeric and dimeric form. Arch Biochem Biophys. 1985 Jul;240(1):102–116. [PubMed]
  • Wang C, Smith RL. Lowry determination of protein in the presence of Triton X-100. Anal Biochem. 1975 Feb;63(2):414–417. [PubMed]
  • van Gelder BF. On cytochrome c oxidase. I. The extinction coefficients of cytochrome a and cytochrome a3. Biochim Biophys Acta. 1966 Apr 12;118(1):36–46. [PubMed]
  • van GELDER B, SLATER EC. The extinction coefficient of cytochrome c. Biochim Biophys Acta. 1962 Apr 23;58:593–595. [PubMed]
  • CONRAD H, SMITH L. A study of the kinetics of the oxidation of cytochrome c by cytochrome c oxidase. Arch Biochem Biophys. 1956 Aug;63(2):403–413. [PubMed]
  • Smith L, Davies HC, Nava M. Oxidation and reduction of soluble cytochrome c by membrane-bound oxidase and reductase systems. J Biol Chem. 1974 May 10;249(9):2904–2910. [PubMed]
  • Smith HT, Ahmed AJ, Millett F. Electrostatic interaction of cytochrome c with cytochrome c1 and cytochrome oxidase. J Biol Chem. 1981 May 25;256(10):4984–4990. [PubMed]
  • Kang CH, Brautigan DL, Osheroff N, Margoliash E. Definitaion of cytochrome c binding domains by chemical modification. Reaction of carboxydinitrophenyl- and trinitrophenyl-cytochromes c with baker's yeast cytochrome c peroxidase. J Biol Chem. 1978 Sep 25;253(18):6502–6510. [PubMed]
  • Ferguson-Miller S, Brautigan DL, Margoliash E. Correlation of the kinetics of electron transfer activity of various eukaryotic cytochromes c with binding to mitochondrial cytochrome c oxidase. J Biol Chem. 1976 Feb 25;251(4):1104–1115. [PubMed]
  • Speck SH, Dye D, Margoliash E. Single catalytic site model for the oxidation of ferrocytochrome c by mitochondrial cytochrome c oxidase. Proc Natl Acad Sci U S A. 1984 Jan;81(2):347–351. [PMC free article] [PubMed]

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