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Kinetics of Accumulation of Ribulose-15-bisphosphate Carboxylase during Greening in Euglena gracilis

机译:裸眼草绿化过程中15-双磷酸核糖15-二磷酸核糖羧化酶积累的动力学

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摘要

The preparation of a rabbit antibody to ribulose-1,5-bisphosphate carboxylase (RuBPCase) from Euglena gracilis and its use to quantitate RuBPCase in dark- and light-grown cells and during light-induced chloroplast development (greening) are described. Light-grown Euglena have at least 36 times more RuBPCase than dark-grown Euglena. Light is required for both the initiation and continued increase in net synthesis of RuBPCase over the dark level: brief illumination 12 hours before exposure to continuous light eliminates the lags in the accumulation and increase in activity of RuBPCase (as well as in chlorophyll accumulation); net synthesis is blocked in greening cells returned to the dark or exposed to 3-(3,4-dichlorophenyl)-1,1-dimethylurea. Streptomycin or cycloheximide prevents RuBPCase accumulation when added at the beginning of greening but only partially blocks accumulation when added after 25 hours of greening. After 24 hours of greening, the activity of RuBPCase per milligram chlorophyll continues to increase slowly while concentration of the enzyme per milligram chlorophyll remains constant. This increased activity may be due to activation of the enzyme as well as to net synthesis.
机译:描述了从裸藻(Euglena gracilis)制备针对核糖1,5-双磷酸羧化酶(RuBPCase)的兔抗体的制备及其在黑暗和光生细胞中以及在光诱导的叶绿体发育过程中(绿化)定量RuBPCase的用途。浅色的Euglena的RuBPCase至少比深色的Euglena多36倍。 RuBPCase的净合成的开始和持续增加都需要光,这是在黑暗条件下进行的:在连续光照射前12小时短暂照明可以消除RuBPCase的积累滞后和活性增加(以及叶绿素积累);净合成被阻止在绿化的细胞中恢复到黑暗或暴露于3-(3,4-二氯苯基)-1,1-二甲基脲。在绿化开始时添加链霉素或环己酰亚胺可防止RuBPCase积累,但在绿化25小时后添加时,链霉素或环己酰亚胺可阻止RuBPCase积累。绿化24小时后,每毫克叶绿素的RuBPCase活性继续缓慢增加,而每毫克叶绿素的酶浓度保持恒定。这种增加的活性可能是由于酶的活化以及净合成所致。

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