Abstract
Two different NAD(P)H dehydrogenases could be demonstrated in the blue-green alga, Aphanocapsa. Both function as quinone reductases using benzoquinone as electron acceptor. One, which was found in the soluble fraction, was NADH specific and showed high sensitivity to rotenone, thenoyltrifluoroacetone and o-phenanthroline. The second dehydrogenase was membrane-bound and used NADH as well as NADPH as substrates. Inhibition by rotenone and o-phenanthroline was less pronounced with the bound enzyme than with the soluble enzyme. Based on studies with NADH or NADPH, the membrane-bound enzyme apparently was associated with a low-temperature EPR signal at g=1.92 in the reduced state, indicative of an iron-sulfur center. The membrane-bound dehydrogenase was solubilized with Triton X-100 and partially purified. This preparation was used for studies of enzyme kinetics and acceptor specificity.
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Abbreviations
- DBMIB:
-
2,5-dibromo methyl isopropylbenzoquinone
- TTFA:
-
thenoyltrifluoroacetone
- E m :
-
midpoint redox potential
References
Bergsma J, Strijker R, Alkema JYE, Seijen HG, Konings WN (1981) Purification and characterization of NADH dehydrogenases from Bacillus subtilis. Eur J Biochem 120:599–606
Biggins J (1969) Respiration in blue-green algae. J Bacteriol 99:570–575
Binder A (1982) Respiration and photosynthesis in energy-transducing membranes of cyanobacteria. J Bioenerg Biomembr 14:271–286
Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254
Cammack R, Luijk LJ, Maguire JJ, Packer L (1970) EPR spectra of photosystem I and other iron protein components in intact cells of cyanobacteria. Biochim Biophys Acta 548:267–275
Godde D (1982) Evidence for a membrane bound NADH-plastoquinone-oxidoreductase in Chlamydomonas reinhardii CW-15. Arch Microbiol 131:197–202
Horton AA (1968) NADH oxidase in blue-green algae. Biochem Biophys Res Commun 32:839–845
Houchins JP, Hind G (1982) Pyridine nucleotides and H2 as electron donors to the respiratory and photosynthetic electron-transfer chains and to nitrogenase in Anabaeno heterocysts. Biochim Biophys Acta 682:86–96
Ingledew WJ, Ohnishi T (1977) The probable site of action of thenoyltrifluoroacetone on the respiratory chain. Biochem J 164:617–620
Ingledew WJ, Prince RC (1977) Thermodynamic resolution of the ironsulfur centers of succinic dehydrogenase of Rhodopseudomonas sphaeroides. Arch Biochem Biophys 178:303–307
Jaworowski A, Mayo G, Shaw DC, Campbell HD, Young IA (1981) Characterization of the respiratory NADH dehydrogenase of Escherichia coli and reconstitution of NADH oxidase in NDH mutant membrane vesicles. Biochemistry 20:3621–3628
MacKinney G (1940) Absorption of light by chlorophyll solutions. J Biol Chem 140:315–322
Malkin R, Chain RK, Kraichoke S, Knaff DB (1981) Studies of the function of the membrane-bound iron-sulfur centers of the photosynthetic bacterium Chromatium vinosum. Biochim Biophys Acta 637:88–95
Myers J, Graham JR, Wang RT (1980) Light harvesting in Anacystis nidulans studied in pigment mutants. Plant Physiol 66:1144–1149
Ohnishi T (1973) Mechanisms of electron transport and energy conservation in the site I region of the respiratory chain. Biochim Biophys Acta 301:105–128
Peschek GA (1980) Electron transport reactions in respiratory particles of hydrogenase-induced Anacystis nidulans. Arch Microbiol 125:123–137
Prince RC, Crowder MS, Bearden AJ (1980) The orientation of the magnetic axes of the membrane-bound iron-sulfur clusters of spinach chloroplasts. Biochim Biophys Acta 592:323–337
Ragan CI (1976) NADH-ubiquinone oxidoreductase. Biochim Biophys Acta 456:249–290
Ragan CI, Racker EI (1973) Resolution and reconstitution of the mitochondrial electron transport chain. J Biol Chem 248:6876–6884
Sandmann G, Malkin R (1983) NADH and NADPH as electron donors to respiratory and photosynthetic electron transport in the blue-green alga Aphanocapsa. Biochim Biophys Acta (submitted)
Scholes RB, Smith L (1968) Composition and properties of the membrane-bound respiratory chain system of Micrococcus dentrificans. Biochim Biophys Acta 153:363–375
Smith AJ, London J, Stanier RY (1967) Biochemical basis of obligate autotrophy in blue-green algae and thiobacilli. J Bacteriol 94:972–983
Thomson JW, Shapiro BM (1981) The respiratory chain NADH dehydrogenase of Escherichia coli. J Biol Chem 256:3077–3084
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Sandmann, G., Malkin, R. NADH and NADPH dehydrogenases from the blue-green alga, Aphanocapsa . Arch. Microbiol. 136, 49–53 (1983). https://doi.org/10.1007/BF00415609
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DOI: https://doi.org/10.1007/BF00415609