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Amino acid synthesis in photosynthesizing spinach spinacia oleracea cells effects of ammonia on pool sizes and rates of labeling from carbon 14 labeled carbon di oxide



Amino acid synthesis in photosynthesizing spinach spinacia oleracea cells effects of ammonia on pool sizes and rates of labeling from carbon 14 labeled carbon di oxide



Plant Physiology 68(2): 292-299



Isolated cells from leaves of S. oleracea were maintained in a state capable of high rates of photosynthetic CO2 fixation for more than 60 h. The incorporation of 14CO2 under saturating CO2 conditions into carbohydrates, carboxylic acids and amino acids, and the effect of NH4 on this incorporation were studied. Total incorporation, specific radioactivity and pool size were determined as a function of time for most of the protein amino acids and for GABA. The measurements of specific radio-activities and of the approaches to 14C saturation of some amino acids indicate the presence and relative sizes of metabolically active and passive pools of these amino acids. Added NH4 decreased C fixation into carbohydrates and increased fixation into carboxylic acids and amino acids. Different amino acids were affected in different and highly specific ways. NH4 caused large stimulatory effects in incorporation of 14C into glutamine (a factor of 21), aspartate, asparagine, valine, alanine, arginine and histidine. No effect or slight decreases were seen in glycine, serine, phenylalanine and tyrosine labeling. In the case of glutamate, 14C labeling decreased, but specific radioactivity increased. The production of labeled GABA was virtually stopped by NH4. Added NH4 apparently stimulates the reactions mediated by pyruvate kinase and phosphoenolpyruvate carboxylase, as seen with other plant systems. The data on the effects of added NH4 on total labeling, pool sizes and specific radioactivities of several amino acids provides a number of indications about the intracellular sites of principal synthesis from C skeletons of these amino acids and the selective nature of effects of increased intracellular NH4 concentration on such synthesis.

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