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Characterization of a covalently linked complex involving ferredoxin and ferredoxin: NADP reductase

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Abstract

Ferredoxin and the flavoprotein, ferredoxin: NADP reductase, have been covalently linked by incubation in the presence of a water soluble carbodiimide. The cross-linking reaction yields an adduct having a 1:1 stoichiometry. The adduct has depressed levels of diaphorase and NADPH oxidase activity and is inactive in reduction of cytochrome c using NADPH as an electron donor. Thus, although similar to an adduct described by Zanetti and coworkers [J Biol Chem 259: 6153–6157 (1984)] in its stoichiometry, the adduct described herein has significantly different enzymatic properties. It is suggested that this may be a reflection of differences in the interaction between the two proteins resulting from differences in experimental conditions in which the two adducts were prepared.

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Abbreviations

Fd:

ferredoxin

Fp:

ferredoxin: NADP reductase

Fd:

Fp covalently linked Fd-Fp adduct

Fd:Fp:

noncovalently linked complex between Fd and Fp

EDC:

1-ethyl-3-(dimethylaminopropyl) carbodiimide

Tris:

tris-hydroxymethylaminomethane

MOPS:

3-(N-morpholino)propane sulfonic acid

DCIP:

2,6-dichloropenolindophenol

References

  1. BatieC and KaminH (1981) The relation of pH and oxidation reduction potential to the association state of the ferredoxin-ferredoxin: NADP reductase complex. J Biol Chem 256: 7756–7763

    Google Scholar 

  2. BatieC and KaminH (1984) Ferredoxin: NADP oxidoreductase, equilibria in binary and ternary complexes with NADP and ferredoxin. J Biol Chem 259: 8832–8839

    Google Scholar 

  3. DavisDJ and San PietroA (1977a) Interactions between spinach ferredoxin and other electron carriers. Arch Biochem Biophys 182: 266–272

    Google Scholar 

  4. DavisDJ and San PietroA (1977b) Evidence for the role of sulfhydryl groups in a pH dependent transition of ferredoxin: NADP oxidoreductase. Arch Biochem Biophys 184: 197–203

    Google Scholar 

  5. Davis DJ (1981) Relationship between substrate binding sites on ferredoxin: NADP reductase. Proc 5th Intl Cong Photosynthesis (ed. G Akoyunoglou) 2: 39–46

  6. DavisDJ and HoughK (1983) Preparation of a covalently linked adduct between plastocyanin and cytochrome f. Biochem Biophys Res Commun 116: 1000–1006

    Google Scholar 

  7. Forti G, Cappelletti A, Nobili RL, Garlaschi FM, Gerola PD and Jennings RC 91983) Interaction of ferredoxin and ferredoxin: NADP reductase with thylakoids. Arch Biochem BViophys 221: 507–513

  8. FortiG and BracaleM (1984) Ferredoxin-ferredoxin: NADP reductase interaction, catalytic differences between soluble and thylakoid bound complex. FEBS Lett 166: 81–84

    Google Scholar 

  9. FoustG, MayhewS and MasseyV (1969) Complex formation between ferredoxin: TPN reductase and electron transfer proteins. J Biol Chem 244: 964–970

    Google Scholar 

  10. GerenLM, StonehuernerJ, DavisDJ and MillettFS (1983) The use of a water-soluble carbodiimide to cross-link cytochrome c to plastocyanin. Biochim Biophys Acta 724: 62–68

    Google Scholar 

  11. GerenLM, O'BrienP, StonehuernerJ and MillettFS (1984) identification of specific carboxylate groups on adrenodoxin that are involved in the interaction with adrenodoxin reductase. J Biol Chem 259: 2155–2160

    Google Scholar 

  12. GozzerC, ZanettiG, GallianoM, SacchiGA, MinchiottiL and CurtiB (1977) molecular heterogeneity of ferredoxin: NADP reductase from spinach leaves. Biochim Biophys Acta 485: 278–290

    Google Scholar 

  13. HackettCS and StrittmatterP (1984) Covalent cross-linking of the active sites of vesicle bound cytochrome b5 and NADH: cytochrome b5 reductase. J Biol Chem 259: 3275–3282

    Google Scholar 

  14. MillettFS, Darley-UsmarV and CapaldiR (1982) Cytochrome c is cross-linked to subunit II of cytochrome c oxidase by a water soluble carbodiimide. Biochemistry 21: 3857–3862

    Google Scholar 

  15. NakamuraS (1970) Initiation of sulfite oxidation by ferredoxin: NADP reductase and ferredoxin system: a model experiment on the superoxide anion radical production by metalloflavoproteins. Biochem Biophys Res Commun 41: 177–183.

    Google Scholar 

  16. NakamuraS and KimuraT (1971) Studies on spinach ferredoxin: NADP reductase: Kinetic studies on the interaction of the reductase and ferredoxin and a possible regulation by ionic strength. J Biol Chem 246: 6235–6241

    Google Scholar 

  17. NakamuraS and KimuraM (1972) Studies on aggregated multienzyme systems: Stimulation of Oxygen Uptake of ferredoxin: NADP reductase-ferredoxin complex by cytochrome c. J Biol Chem 247: 6462–6468.

    Google Scholar 

  18. NelsonN and NeumanJ (1969a) Interaction between ferredoxin and ferredoxin: NADP reductase in pyridine nucleotide photoreduction and some partial reactions I. J Biol Chem 244: 1926–1931

    Google Scholar 

  19. NelsonN and NeumanJ (1969b) Interaction between ferredoxin and ferredoxin: NADP reductase in pyridine nucleotide photoreduction and some partial reactions II. J Biol Chem 244: 1932–1936

    Google Scholar 

  20. PeteringDH and PalmerG (1970) Properties of spinach ferredoxin in anaerobic urea solutions: comparison with the native protein. Arch Biochem Biophys 141: 456–464

    Google Scholar 

  21. ShinM (1968) Ferredoxin-NADP reductase. In: YagiK (ed) Flavins and Flavoproteins pp 1–5 Univ Tokyo Press Tokyo Japan

    Google Scholar 

  22. ShinM and SanPietro A (1968) Complex formation of ferredoxin: NADP reductase with ferredoxin and NADP. Biochem Biophys Res Commun 33: 33–42

    Google Scholar 

  23. VieiraBJ and DavisDJ (1983) Chemical modification of spinach ferredoxin with diethylpyrocarbonate: evidence for the essential nature of the histidyl residue. Biochem Biophys Res Commun 112: 508–514

    Google Scholar 

  24. VieiraBJ and DavisDJ (1985) Chemical modification of carboxyl groups on spinach ferredoxin alters its ability to interact with ferredoxin: NADP reductase. Biochem Biophys Res Commun 129: 467–471

    Google Scholar 

  25. VieiraBJ and DavisDJ (1986) Interaction of ferredoxin with ferredoxin: NADP reductase: effects of chemical modification of ferredoxin. Arch Biochem Biophys 247: 140–146

    Google Scholar 

  26. VieiraBJ, ColvertK and DavisDJ (1986) Chemical modification and cross-linking as probes of regions on ferredoxin involved in its interaction with ferredoxin: NADP reductase. Biochim Biohys Acta 851: 109–122

    Google Scholar 

  27. WaldmeyerB, BechtoldR, BosshardHR and PoulosT (1982) The cytochrome c peroxidase-cytochrome c electron transfer complex: experimental evidence for a hypothetical model. J Biol Chem 257: 3073–3076

    Google Scholar 

  28. WeberK and OsbornM (1969) The reliability of molecular weight determinations by dodecyl sulfate polyacrylamide gel electrophoresis. J Biol Chem 244: 4406–4412

    Google Scholar 

  29. ZanettiG, AlivertiA and CurtiB (1984) A cross-linked complex between ferredoxin and ferredoxin: NADP reductase. J Biol Chem 259: 6253–6257

    Google Scholar 

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Author notes

  1. Kim K. Colvert

    Present address: Department of Chemistry, Southwest Missouri State University, 65802, Springfield, MO, USA

Authors and Affiliations

  1. Department of Chemistry and Biochemistry, University of Arkansas, 72701, Fayetteville, AR, USA

    Kim K. Colvert & Danny J. Davis

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  1. Kim K. Colvert
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  2. Danny J. Davis
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Colvert, K.K., Davis, D.J. Characterization of a covalently linked complex involving ferredoxin and ferredoxin: NADP reductase. Photosynth Res 17, 231–245 (1988). https://doi.org/10.1007/BF00035450

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  • DOI: https://doi.org/10.1007/BF00035450

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