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  • 1
    ISSN: 1432-2048
    Keywords: Avena ; Chlorophyll biosynthesis ; Etioplasts ; Geranylgeranyldiphosphate ; Protoplasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The uptake of [1-3H]geranylgeranyl diphosphate (GGPP) into protoplasts and intact etioplasts and the metabolic interconversion therein was studied after a 2 min pulse of white light. The chlorophyll synthetase reaction, Chlide+GGPP→ChlGG, was taken as a natural probe for the etioplast compartment. This reaction yields labeled ChLGG and, by hydrogenation, labeled ChlP, when [1-3H]GGPP receives access to the etioplast stroma. It was found that penetration across the plastid envelope was rapid and that penetration across the plasma membrane of protoplasts, however, was slow. A cellular pool of soluble GGPP was detected. This pool was lost, in part, during preparation of the protoplasts and almost completely during preparation of the etioplasts. The membrane-bound phytol pool of etioplasts could not be replaced by exogenous [3H]GG. The endogenous GG and phytol pools of protoplasts, which were larger than those of etioplasts, could be replaced in part by exogenous [3H]GGPP. That part of this pool exists as soluble GGPP or as a direct precursor in the cytoplasm is discussed.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2048
    Keywords: Avena ; Etioplasts ; Mitochondria ; Phytochrome
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Phytochrome was determined in etiolated laminae of Avena sativaL. either without pretreatment or after 5 min of red irradiation followed by different periods of darkness (0–24 h). At given intervals laminae were homogenized and phytochrome was determined spectrophotometrically in the total homogenate and in purified etioplasts and mitochondria. Enhanced specific activity of phytochrome was found in all fractions after the irradiation in comparison to dark controls. Phytochrome destruction was observed in all fractions at the beginning of the subsequent dark period. Whereas the homogenate and the mitochondrial fraction showed a continuous destruction so that phytochrome reached a level far below that in etiolated plants, the phytochrome level in the plastid fraction reacheda minimum at 2 h with a subsequent increase beyond the dark level. This increase was most pronounced between 4 and 8 h after the red irradiation. The results are discussed in terms of the destruction and possible de novo synthesis of phytochrome that may be different in mitochondria and plastids.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-2048
    Keywords: Avena ; Calmodulin ; Etiolated seedling ; Phytochrome ; Protein kinase ; (ATP-dependent)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A protein-kinase activity which is co-purified with phytochrome from etiolated oat seedlings was investigated in some detail. Whereas phytochrome was always phosphorylated in solution (together with some contaminating protein bands), radioactive phosphate was not found in the phytochrome band after native gel electrophoresis and incubation of the entire gel with labeled ATP. Since protein kinases are usually autophosphorylated under these conditions, the result shows that the kinase activity does not reside in the phytochrome molecule itself. Radioactivity was exclusively detected in a band with the apparent molecular weight 450 kDa; sodium-dodecyl-sulfate gel electrophoresis revealed an apparent molecular weight of 60 kDa for the phosphorylated subunit. The N-terminal amino-acid sequence A L E S A G K Q L V P W was determined for this subunit which is a potential candidate for the protein kinase. The optimum conditions (pH, metal ion concentration) and kinetics of the phosphorylation reaction were determined. The presumed connection between proteinkinase activity and the signal chain leading from the far-red-absorbing form of phytochrome to physiological responses still awaits elucidation.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-2048
    Keywords: Avena ; Photoreceptor ; Phototropism ; Plasma membrane ; Protein kinase ; Protein phosphorylation (blue-light-dependent)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Blue light induced the phosphorylation of a 116-kDa plasma-membrane-associated protein in dark-grown seedlings from Avena sativa L. The response was restricted to the phototropically sensitive tissue of the coleoptile tip. Surprisingly, this protein showed different properties in membrane preparations from plants that were grown for 3 d than in those from 5-d-old seedlings. In contrast to the younger coleoptiles, in 5-d-old seedlings phosphorylation of the 116-kDa protein depended strictly on the addition of Triton X-100 or other non-ionic detergents and was not abolished when the membranes were pretreated with trypsin. These latter membranes were also characterized by the appearance of two additional bluelight-regulated phosphoproteins of slightly lower molecular masses, exhibiting properties similar to the 116-kDa protein from 3-d-old plants. The data, together with solubilization studies, indicate that the 116-kDa protein is strongly membrane-bound only at the very beginning of seedling development and becomes more loosely associated in the course of coleoptile growth. In addition, we demonstrate that the capacity of the light-activated photoreceptor to recover photosensitivity in the dark also can occur under in-vitro conditions.
    Type of Medium: Electronic Resource
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