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Energy Metabolism of Rickettsia typhi: Pools of Adenine Nucleotides and Energy Charge in the Presence and Absence of Glutamate

机译:伤寒立克次体的能量代谢:存在和不存在谷氨酸的腺嘌呤核苷酸池和能量电荷

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摘要

The obligate intracellular bacterium Rickettsia typhi was examined for its ability to generate and maintain an adenylate energy charge in an extracellular environment. Freshly purified organisms were incubated, at 34°C and pH 7.4, with or without glutamate and various other metabolites, and the levels of ATP, ADP, and AMP were determined. Of the metabolites tested, glutamate and glutamine were the most effective for the generation of ATP. In the presence of glutamate, there was a rapid increase in the level of ATP, followed by a moderate decrease during 150 min of incubation. The energy charge increased from a level of 0.2 to 0.5 to about 0.7 to 0.75, and then slowly declined to about 0.45 to 0.6. In the absence of glutamate, after an occasional initial surge in ATP level as the temperature was changed from 4 to 34°C, there was a sharp decline in both ATP and energy charge (to 0.1 and sometimes to 0.01). The rickettsiae maintained their ability to regenerate their energy charge upon the addition of glutamate for about 30 min, but this ability declined with further incubation. In contrast to Escherichia coli, the decline in ATP in R. typhi was accompanied by a sharp increase in the level of AMP and the total adenylate pool. No adenine or adenosine was recovered from rickettsiae incubated with labeled AMP, ADP, or ATP. From these experiments and the demonstration reported elsewhere that rickettsiae transport the adenine nucleotides, it can be concluded that the adenylate energy charge in R. typhi is governed by the salvage of the adenine nucleotides rather than their unphosphorylated precursors. Thus, R. typhi undergoes greater shifts in energy charge than other bacteria, a phenomenon which may account for their instability in an extracellular environment. Under optimal conditions the adenylate energy charge of R. typhi approaches levels that border on those generally regarded as adequate for growth.
机译:检查了专性细胞内细菌伤寒立克次体在细胞外环境中产生和维持腺苷酸能量电荷的能力。将新鲜纯化的生物体在有或没有谷氨酸和各种其他代谢物的条件下于34°C和pH 7.4孵育,并测定ATP,ADP和AMP的水平。在测试的代谢产物中,谷氨酸和谷氨酰胺最有效地产生了ATP。在存在谷氨酸的情况下,ATP水平迅速增加,随后在孵育150分钟期间适度降低。能量电荷从0.2到0.5的水平增加到大约0.7到0.75,然后缓慢下降到大约0.45到0.6。在没有谷氨酸的情况下,随着温度从4到34°C的变化,ATP偶尔出现最初的激增后,ATP和能量电荷都急剧下降(下降到0.1,有时下降到0.01)。加入谷氨酸盐约30分钟后,立克次体保持了其再生能量电荷的能力,但是这种能力随着进一步温育而下降。与大肠杆菌相反,伤寒沙门氏菌中ATP的下降伴随着AMP水平和总腺苷酸池的急剧增加。从与标记的AMP,ADP或ATP孵育的立克次体中未回收腺嘌呤或腺苷。从这些实验和在其他地方报道的示范立克次体转运腺嘌呤核苷酸,可以得出结论,伤寒沙门氏菌中腺苷酸的能量电荷是由腺嘌呤核苷酸的抢救而不是其未磷酸化的前体决定的。因此,伤寒沙门氏菌比其他细菌经历更大的能量转移,这种现象可能解释了它们在细胞外环境中的不稳定性。在最佳条件下,伤寒沙门氏菌的腺苷酸能电荷接近通常被认为适合生长的水平。

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