Abstract
Hemoproteins were localized in the cyanobacteriumAnabaena cylindrica with diaminobenzidine (DAB). Incubation of whole cells in the light with DAB resulted in deposition of oxidized DAB on the lamellae of the vegetative cells and central heterocyst region. This reaction was greatest at pH 7.5, light-dependent, insensitive to 3-(3,4-dichlorophenyl)-1, 1-dimethyl urea, and abolished by glutaraldehyde fixation. A light-independent oxidation of DAB was also observed with light and electron microscopy in the “honeycomb” region and periphery of heterocysts. This reaction was greatest at pH 7.5, enhanced by H2O2, and active in glutaraldehyde-fixed frozen sections. Inhibitors such as sodium cyanide, sulfide, and hydroxylamine severely reduced DAB oxidation and nitrogenase activity under aerobic but not anaerobic conditions. These results indicate that the heme proteins, localized in heterocysts by light-independent DAB oxidation, are involved in the oxygen-protection mechanism of the O2-labile nitrogenase.
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Literature Cited
Allen, M. B., Arnon, D. I. 1955. Studies on nitrogen-fixing blue-green algae I. Growth and nitrogen fixation byAnabaena cylindrica Lemm. Plant Physiology30:366–372.
Bottomley, P., Stewart, W. D. P. 1977. The measurement and significance of ATP pools in filamentous blue-green algae. British Physological Journal11:69–82.
Cammer, W., Moore, C. L. 1973. Oxidation of 3,3′-diaminobenzidine by rat liver mitochondria. Biochemistry12:2502–2509.
Chua, N. H. 1972. Photooxidation of 3,3′-diaminobenzidine by blue-green algae andChlamydomonas reinhardii. Biochimica et Biophysica Acta267:179–189.
Essner, E. 1974. Hemoproteins, pp. 1–33. In: Hayat, M. A. (ed.), Electron microscopy of enzymes. Principles and methods, vol. 2. New York: Van Nostrand Reinhold.
Fahimi, H. D. 1970. The fine structural localization of endogenous and exogenous peroxidase activity in Kupffer cells of rat liver. Journal of Cell Biology47:247–262.
Fay, P., Walsby, A. E. 1966. Metabolic activities of isolated heterocysts of the blue-green algaAnabaena cylindrica. Nature209:94–95.
Frederick, S. E., Newcomb, E. H. 1969. Cytochemical localization of catalase in leaf microbodies. Journal of Cell Biology43:343–353.
Hallier, U. W., Park, R. B. 1969. Photosynthetic light reactions in chemically fixed spinach thylakoids. Plant Physiology44:544–546.
Herzog, V., Miller, F. 1972. Endogenous peroxidase in the lacrimal gland of the rat and its differentiation against injected catalase and horseradish-peroxidase. Histochemie30:235–246.
Jurtshuk, P., Jr., McQuitty, D. N., Riley, W. H., IV. 1979. Use of 3,3′-diaminobenzidine as a biochemical electron donor for studies on terminal cytochrome oxidase activity inAzotobacter vinelandii. Current Microbiology2:349–354.
Lang, N. J., Fay, P. 1971. The heterocysts of blue-green algae II. Details of ultrastructure. Proceedings of the Royal Society, London B,178:193–203.
Lauritis, J. A., Vigil, E. L., Sherman, L., Swift, H. 1975. Photosynthetically-linked oxidation of diaminobenzidine in blue-green algae. Journal of Ultrastructure Research53:331–344.
Murry, M. A., Benemann, J. R. 1979. Nitrogenase regulation inAnabaena cylindrica. Plant and Cell Physiology20:1391–1401.
Nir, I., Pease, D. C. 1973. Chloroplast organization and the ultrastructural localization of photosystems I and II. Journal of Ultrastructure Research42:534–550.
Nir, I., Seligman, A. M. 1970. Photooxidation of diaminobenzidine (DAB) by chloroplast lamellae. Journal of Cell Biology46:617–620.
Novikoff, A. G., Goldfischer, S. 1969. Visualization of peroxisomes (microbodies) and mitochrondria with diaminobenzidine. Journal of Histochemistry and Cytochemistry17:675–680.
Patterson, C. O. P., Myers, J. 1973. Photosynthetic production of hydrogen peroxide byAnacystis nidulans. Plant Physiology51:104–109.
Peterson, R. B., Burris, R. A. 1978. Hydrogen metabolism in isolated heterocysts ofAnabaena 7120. Archives of Microbiology116:125–132.
Poux, N. 1972. Localisation d'activités enzymatiques dans le méristéme radiculaire deCucumis sativus L. IV. Réactions avec la diaminobenzidine mes evidence de peroxysomes. Journal de Microscopie (Paris)14:183–218.
Roels, F. 1973. Le 2,6-dichlorophénolindophénol est-il un inhibiteur spécifique de la catalase? Comptes Rendus de l'Academie des Sciences (Paris) Serie D277:889–891.
Thomas, J. 1970. Absence of the pigments of photosystem, II of photosynthesis in heterocysts of a blue-green alga. Nature (London)228:181–183.
Van Baalen, C. 1965. Quantitative surface plating of coccoid blue-green algae. Journal of Phycology1:19–22.
Vigil, E. L. 1973. Structure and function of plant microbodies. Subcellular Biochemistry2:237–285.
Wahal, C. K., Bhattacharya, N. C., Talpasyi, E. R. S. 1974. Study of some isoenzyme patterns ofAnabaena ambiqua with and without heterocysts. Biochemie und Physiologie der Pflanzen165:351–361.
Weare, N. M., Benemann, J. R. 1973. Nitrogen fixation byAnabaena cylindrica. I. Localization of nitrogen fixation in the heterocysts. Archives of Microbiology90:323–332.
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Murry, M.A., Olafsen, A.G. & Benemann, J.R. Oxidation of diaminobenzidine in the heterocysts ofAnabaena cylindrica . Current Microbiology 6, 201–206 (1981). https://doi.org/10.1007/BF01566972
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DOI: https://doi.org/10.1007/BF01566972