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Preparation of a highly active ATPase of the mesophilic cyanobacterium Spirulina maxima

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Abstract

In this work, we report new studies on the ATPase attached to the photosynthetic membranes of the mesophilic cyanobacterium Spirulina maxima. This enzyme does not display persistent latency as had been previously reported for the ATPase of Spirulina platensis. The enzyme is readily activated by the careful application of methods currently used to activate chloroplast CF1. Photosynthetic membranes of Spirulina maxima show a Mg2+-dependent ATPase activity of 195±25 μmol Pi (mg chl)−1 h−1 after a light plus dithiothreitol (DDT) treatment. Methanol treatment of these membranes elicits Mg2+-dependent ATPase activity of 222±18 μmol Pi (mg chl)−1 h−1.

Here, we also describe the purification of the soluble coupling factor AF1 of Spirulina maxima. This enzyme is unique among mesophilic cyanobacterial F1 preparations in regard to its high specific Ca2+-dependent ATPase activity after heat treatment (14.75±1.91 μmol Pi (mg prot)−1 min−1) and its room temperature stability.

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Abbreviations

AF1 :

cyanobacterial coupling factor

DTT:

dithiothreitol

DEAE cellulose:

diethylaminoethyl cellulose

DCCD:

N,N'-dicyclohexylcarbodiimide

EDTA:

ethylenediamine tetracetic, sodium salt

PAGE:

polyacrylamide gel electrophoresis

PMS:

phenazine methosulfate

PMSF:

phenylmethylsulfonyl fluoride

MV:

methylviologen

SDS:

sodium dodecylsulfate

TPSnCl:

triphenyltin chloride

Tris:

tris (hydroxymethyl) aminomethane

tricine:

N Tris (hydroxymethyl) methylglycine

BAEE:

Nα-benzoyl-L-arginine ethyl ester

References

  1. Anthon G and Jagendorf A (1983) Effect of methanol on spinach thylakoid ATPase. Biochim Biophys Acta 723: 358–365

    Google Scholar 

  2. Andrews A (1981) Electrophoresis. Oxford: Claren press

    Google Scholar 

  3. Arnon D (1949) Copper enzymes in isolated chloroplasts. Polyphenol oxidase in Beta vulgaris. Plant Physiol 24: 1–15

    Google Scholar 

  4. Bates F, Phelps F and Snyder C (1927) Saccarimetry, the properties of commercial sugars and their solutions. In “International Critical Tables” vol II pp. 334–355 New York: McGraw Hill

    Google Scholar 

  5. Beechey R, Hubbard S, Linnet P, Mitchell D and Munn E (1975) A simple and rapid method for the preparation of adenosine triphosphate from submitochondrial particles. Biochem J 148: 533–537

    Google Scholar 

  6. Bensadoun A and Weinstein D (1976) Assay of proteins in the presence of interfering materials. Annal Biochem 70: 241–250

    Google Scholar 

  7. Binder A, Jagendorf A and Ngo E (1978) Isolation and composition of the subunits of spinach chloroplast coupling factor protein. J Biol Chem 253: 3094–3100

    Google Scholar 

  8. Binder A and Bachofen R (1979) Isolation and characterization of a coupling factor I ATPase of the thermophilic blue green algae Mastigocladus laminosus FEBS Lett 104: 66–70

    Google Scholar 

  9. Binder A, Bohler M, Wolf M and Bachofen R (1981) The proton ATPase of the thermophilic cyanobacterium Mastigocladus laminosus. In Akoyunoglou G ed. Proceedings of the 5th International Photosynthesis Congress Vol 6 pp 771–778. Philadelphia: Balaban International Science Services

    Google Scholar 

  10. Bradford M (1976) A rapid and senstive method for quantitation of microgram quantities of protein utilizing the principle of protein dye binding. Annal Biochem 72: 248–254

    Google Scholar 

  11. Deters D, Racker E, Nelson N and Nelson H (1975) Partial resolution of the enzymes catalyzing photophosphorylation XV Approaches to the active site of coupling factor I. J Biol Chem 250: 1041–1047

    Google Scholar 

  12. Hachimori A, Muramatsu N & Nasch Y (1970) Studies on an ATPase of thermophilic bacteria. I. Purification and properties. Biochim Biophys Acta 206: 426–437

    Google Scholar 

  13. Laemmli U (1970) Cleavage of structural protein during the assembly of the head of bateriophage T4. Nature 227: 680–685

    Google Scholar 

  14. Lerma C and Gómez-Lojero C (1982) Photosynthetic Phosphorylation of a membrane preparation of the cyanobacterium Spirulina maxima. Biochim Biophys Acta 680: 181–186

    Google Scholar 

  15. Lien S and Racker E (1971) Preparation and assay of chloroplast coupling factor CF1 in San Pietro A ed. “Methods in Enzymology” Vol 23 Photosynthesis. Part A pp 547–555 New York: Academic Press

    Google Scholar 

  16. Lubberding H, Offerijns F, Willemien A and Vel deVries J (1981) Characterization of ATPase of the thermophilic cyanobacterium Synechococcus lividus. In Akoyunoglou G ed “The Proceedings of the 5th International Congress of Photosynthesis” Vol 6 pp 779–788 Philadelphia: Balaban International Science Services

    Google Scholar 

  17. McCarty R and Racker E (1968) Partial resolution of the enzymes catalyzing photophosphorylation. III Activation of adenosine triphosphatase and 32P-labeled ortophosphate adenosine triphosphate exchange in chloroplasts. J Biol Chem 243: 129–137

    Google Scholar 

  18. McCarty R (1979) Roles of a coupling factor for photophosphorylation in chloroplasts. Ann Rev Plant Physiol 30: 79–104

    Google Scholar 

  19. Melandri B and Baccarini-Melandri A (1976) Coupling factor ATPases from photosynthetic bacteria. J Bioenerg 8: 109–119

    Google Scholar 

  20. Nishimura M, Ito T and Chance B (1962) Studies on Bacterial photophosphorylation. Biochim Biophys Acta 59: 177–183

    Google Scholar 

  21. Ortiz F, Acosta A and Gómez-Puyou A (1982) Characteristics of Adenylyl imidophosphate and ADP binding sites in soluble and particulate mitochondrial ATPase. Biochim Biophys Acta 679: 466–473

    Google Scholar 

  22. Owers-Narhi L, Robinson S, Selvius deRoo C and Yocum C (1979) Reconstitution of cyanobacterial photophosphorylation by a latent Ca2+-ATPase. Biochem Biophys Res Commun 90: 1025–1031

    Google Scholar 

  23. Penefsky H (1974) Mitochondrial and chloroplast ATPase. In Boyer P ed. “The enzymes” Vol X pp 375–394. New York: Academic Press

    Google Scholar 

  24. Sakurai H, Shinohara K, Hisabori T and Shinihara K (1981) Enhancement of adenosine triphosphatase activity of purified chloroplast coupling factor I in an aqueous organic solvent. J Biochem 90: 95–102

    Google Scholar 

  25. Schatz G, Penefsky H and Racker E (1967) Partial resolutions of the enzymes catalyzing oxidative phosphorylation XIV Interaction of purified mitochondrial adenosine triphosphatase from baker's yeast with submitochondrial particles from beef heart. J Biol Chem 242: 2552–2560

    Google Scholar 

  26. Schuster S (1979) Effect of organic solvents on the beef heart mitochondrial adenosine triphosphatase. Biochemistry 18: 1162–1167

    Google Scholar 

  27. Summer J. (1944) A method for the colorimetric determination of phosphorus. Science 100: 413–414

    Google Scholar 

  28. Trebst A (1974) Energy conservation in photosynthetic electron transport of chloroplasts. Ann Rev Plant Physiol 25: 423–458

    Google Scholar 

  29. Vambutas V and Racker E (1965) Partial resolution of the enzymes catalyzing photophosphorylation. I Stimulation of photophosphorylation by a preparation of a latent Ca2+-dependent adenosine triphosphatase from chloroplasts. J Biol Chem 240: 2660–2667

    Google Scholar 

  30. Vogel G and Steinhart R (1976) ATPase of E. coli. Purification, dissociation and reconstitution of the active complex from the isolated subunits. Biochemistry 15: 208–216

    Google Scholar 

  31. Weber K and Osborn M (1969) The reliability of molecular weight determinations by dodecylsulfate-polyacrylamide gel electrophoresis. J Biol Chem 244: 4406–4412

    Google Scholar 

  32. Wolf M Binder A and Bachofen R (1981) Influence of growth temperature on structure thermostability and kinetic properties of the ATPase coupling factor AF1 of a thermophilic blue green (cyanobacterium). Eur J Biochem 118: 423–427

    Google Scholar 

  33. Yoshida M, Sone N, Hirata H and Kagawa Y (1985) A highly stable adenosine triphosphatase from a thermophilic bacterium. J Biol Chem 250: 7910–7916

    Google Scholar 

  34. Younis H, Winget G and Racker E (1977) Requirement of the δ subunit of chloroplast coupling factor I for photophosphorylation. J Biol Chem 252: 1814–1818

    Google Scholar 

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Authors and Affiliations

  1. Departamento de Bioquímica, Centro de Investigación y de Estudios Avanzados del Instituto, Politécnico Nacional, Apartado Postal 14-740, 07000, México 14, D.F., México

    Claudia Lerma & Carlos Gómez-Lojero

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  1. Claudia Lerma
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  2. Carlos Gómez-Lojero
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Lerma, C., Gómez-Lojero, C. Preparation of a highly active ATPase of the mesophilic cyanobacterium Spirulina maxima . Photosynth Res 11, 265–277 (1987). https://doi.org/10.1007/BF00055066

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  • DOI: https://doi.org/10.1007/BF00055066

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