Recovery of plastids from photooxidative damage: Significance of a plastidic factor |
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Authors: | C Schuster R Oelmüller R Bergfeld H Mohr |
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Institution: | (1) Department of Botany, Faculty of Agriculture and Horticulture, Technical University of Munich, D-8050 Freising 12, Federal Republic of Germany |
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Abstract: | Glyoxysomal citrate synthase (gCS) was purified from crude extracts of watermelon (Citrullus vulgaris Schrad.) cotyledons, yielding a homogenous protein with a subunit MW of 48 kDa. The enzyme was selectively inhibited by 5,5-dithiobis-(2-nitrobenzoic acid), allowing quantification in the presence of the mitochondrial isoenzyme (mCS). Differences were also observed with respect to inhibition by ATP (k
i=2.6 mmol · l-1 for gCS, k
i=0.33 mmol · l-1 for mCS). The antibodies prepared against gCS did not cross-react with mCS. The immunocytochemical localization of gCS by the indirect protein A-gold procedure was restricted to the glyoxysomal membrane or the peripheral matrix of glyoxysomes. Other compartments, e.g. the endoplasmic reticulum, were not labeled. Xenopus oocytes were used for the translation of watermelon polyadenylated RNA (poly(A)+RNA). A translation product with a MW of 51 kDa was immunoprecipitated by the anti-gCS antibodies. It was absent in controls without poly(A)+RNA or with preimmune serum. A similar translation product was also immunoprecipitated after cell-free synthesis of watermelon poly(A)+RNA in a reticulocyte system, in contrast to the in-vivo labeled gCS (48 kDa). It was concluded that gCS is synthesized as a higher-molecular-weight precursor.Abbreviations DTNB
5,5-dithiobis-(2-nitrobenzoic acid)
- gCS
glyoxysomal citrate synthase
- gMDH
glyoxysomal malate dehydrogenase
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k
i
inhibitor constant
- mCS
mitochondrial citrate synthase
- OAA
oxaloacetate
- poly(A)+RNA
polyadenylated RNA
- SDS-PAGE
sodium dodecyl sulfate-polyacrylamide gel electrophoresis |
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Keywords: | Citrate synthase (cell-free synthesis) Citrullus (citrate synthase) Cotyledon (citrate synthase) Glyoxysome Xenopus oocytes |
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