A method for extracting intact chloroplast and cytoplasmic ribosomal RNA from leaves

A method is described for extracting intact chloroplast and cytoplasmic ribosomal RNA from leaves of two higher plant species. Sodium dodecyl sulfate (1%) and 25 mM magnesium ions are required to inhibit ribonuclease action during RNA purification by phenol deproteinization. The ethanol-precipitated RNA product, including 23s chloroplast ribosomal RNA, is completely stable during electrophoresis in the absence of magnesium ions, even in the presence of EDTA. The $\text{in}$$\text{vivo}$ mole fraction of chloroplast ribosomes relative to cytoplasmic ribosomes is estimated. Bentonite is shown to cause preferential losses of chloroplast RNA during extraction.     

Ribosome specificity for the formation of guanosine polyphosphates

Ribosomes obtained from $\text{Bacillus brevis}$ (ATCC 8185) were slightly active in synthesizing guanosine polyphosphates, which activity was greatly stimulated by addition of $\text{Escherichia coli}$ stringent factor. $\text{Chlamydomonas reinhardtii}$ chloroplast ribosomes did not produce guanosine polyphosphates on incubation but responded with abundant synthesis to addition of the stringent factor from $\text{E. coli}$. In contrast, cytoplasmic ribosomes from the same organism did not respond. Interchange experiments between either subunit from chloroplasts with the $\text{E. coli}$ counterparts showed good activity. When the small subunit of cytoplasmic $\text{Chlamydomonas}$ ribosomes was combined with the large subunit of $\text{E. coli}$ or of chloroplasts, a small but definite response was obtained.     

Coordination of adenylate energy charge and phosphorylation state during ischemia and under physiological conditions in rat liver and kidney

The relation of the adenylate energy charge $\text{(}\text{ATP}\text{+}\text{1}\text{2}\text{ADP/ATP}\text{+}\text{ADP}\text{+}\text{AMP}\text{)}$ to the phosphorylation state (ATP)/(ADP)(HPO₄^2?) in rat liver and kidney was analyzed. Under physiological conditions and in ischemia, the two regulatory parameters, calculated from reported values for adenine nucleotides and inorganic phosphate (P_i) and from new observations, were closely coordinated. Energy charge was an inverse linear function of P_i and -log (1 - energy charge) was a positive linear function of log phosphorylation state. To evaluate experimental data with known energy charge, but unknown P_i, and to determine the theoretical relation between energy charge and phosphorylation state, P_i was estimated from a) the regression equation: P_i, μmol/g wet wt tissue = 1.05 - energy charge/0.073 and b) the empirical relationship: (P_i/2P_a) + energy charge = k, where P_a = σAMP + 2ADP + 3ATP and k = 1. With both estimates, the relation between phosphorylation state and energy charge for the experimental data was, within error, the same as that observed with measured P_i and concordant with theoretical values. Over the physiological range of energy charge (～0.85 – 0.95, log phosphorylation state ～3.3 – 4.3), apparent ΔG_ATP (×2) was closer to the range of ΔG observed by Wilson $\text{et al}$ (Biochem. J. $\text{140}$:57, 1974) for transfer of two electrons from mitochondrial NAD to the cytochrome c couple than the ΔG_ATP (×2) they reported, supporting their conclusion that near-equilibrium exists between the mitochondrial respiratory chain and the cytoplasmic phosphorylation state under physiological conditions. From evidence presented, it is postulated that the phosphorylation state is regulated by the adenylate energy charge.     

Changes in predominant energy substrate after hepatectomy

The changes in the energy substrate utilized by the remnant liver were studied in relation to the changes in the cellular energy status of 25 and 70% hepatectomized rabbits. In 25% hepatectomized rabbits, the energy charge $\text{(}\text{(}\text{ATP}\text{+0.5}\text{ADP}\text{)}\text{(}\text{ATP}\text{+}\text{ADP}\text{+}\text{AMP}\text{)}\text{)}$ level of the remnant liver remained unchanged, the energy substrate of which was predominantly glucose, rather than fatty acid. In contrast, in 70% hepatectomized rabbits, the energy production by the mitochondria was mainly dependent upon fatty acid oxidation at the early period after hepatectomy when the energy charge level decreased remarkably, and then upon glucose oxidation, concomitant with the restoration of the energy charge. It is suggested that the changes in the energy substrate utilized are closely related to those in the energy charge level and the mitochondrial phosphorylative activity of the remnant liver following hepatectomy.     

Quantum requirements for proton uptake in spinach chloroplasts

Studies of electron and proton transport in chloroplast preparations (Type D) from spinach (Spinacea oleracea L.) yield three basic results. First, in electron transport catalyzed by methyl viologen from water to oxygen at pH 7.6, the quantum requirement for electron transport ($\text{hv}\text{e}{}^{\mathrm{?}}$) was 2.2, while the corresponding requirement for proton transport ($\text{h}\text{v}\text{H}{}^{\mathrm{+}}$) was 1.2. Second, the electron and proton quantum requirements were relatively independent of the individual chloroplast preparation or certain components of the resuspension medium, but did depend upon the reaction medium's initial pH. Third, measurable electron and proton transport did not occur under 715-nm illumination, nor did such activities occur in the presence of DCMU under 645-nm illumination when methyl viologen was used as the electron transport cofactor. These experimental results reconcile the quantum requirement of proton transport with Mitchell's chemiosmotic theory for chloroplast energy transduction and resolve a long standing controversy regarding the quantum requirement in chloroplast thylakoids.     

Light scattering and quenching of 9-aminoacridine fluorescence as indicators of the phosphorylation state of the adenylate system in intact spinach chloroplasts

Upon illumination of suspensions of intact chloroplasts, fluorescence of 9-aminoacridine was quenched, light scattering was increased, chlorophyll fluorescence was decreased after an initial increase, and chloroplast $\text{ATP}\text{ADP}$ ratios were increased. The response of 9-aminoacridine fluorescence quenching and light scattering to light intensity, anaerobiosis and inhibition of electron transport by DCMU was similar to that shown by chloroplast $\text{ATP}\text{ADP}$ ratios. It is discussed under what conditions 9-aminoacridine fluorescence quenching or light scattering can be used to monitor changes in the phosphorylation state of the chloroplast adenylate system.     

Light-dependent development of photosynthetic competence in Scenedesmus mutant no. 8

The heterotrophically grown, $\text{P-700-}\text{free}$ mutant No. 8 of Scenedesmus obliquus is unable to carry out photosynthesis. Yet, chloroplast particles isolated from the alga reduced ferricyanide. They also reduced methyl viologen in the presence of the artificial donor reduced 2,6-dichlorophenol indophenol with a low yield but an appreciable saturation rate. NADP reduction or $\text{P-700}$ turn-over could not be detected.When grown mixotrophically, the mutant showed increasing $\text{P-700}$ activity with a concomitant increase in the rate of photosynthesis. Both activities were lost again when the algae were returned to darkness. Isolated chloroplast particles showed a good $\text{P-700}$ turn-over and reasonable rates of NADP reduction.The data suggest that the mutation occurred at a site preceding the formation of the pigment. The results on the photochemical activities are discussed in the light of reports concerning the involvement of $\text{P-700}$ in linear electron transport.     

The adenylate energy charge in marine phytoplantkon: The effect of temperature on the physiological state of Skeletonema costatum (Grev.) Cleve

The adenylate energy charge $\text{([ATP] +}\text{1}\text{2}\text{[ADP])}\text{[0ATP+ADP+AMP]}$ was measured in axenic batch cultures of Skeletonema costatum (Grev.) Cleve at 2°, 10°, 15°, 20°, 24° and 30°C. The results suggest that this eurythermal diatom is physiologically capable of adapting to the 28 °C range of temperature with little apparent difference in the potential energy available to the cell. In N-limited continuous cultures at 15 °C, the energy charge values were lower than those observed in batch culture by 0.2, implying nutrient stress may result in decreased intracellular chemical energy. The utilization of the adenylate energy charge as an indicator of physiological state is suggested.     

Transport as the basis of the kok effect. Levels of some photosynthetic intermediates and activation of light-regulated enzymes during photosynthesis of chloroplasts and green leaf protoplasts

Experiments were performed with intact chloroplasts and leaf cell protoplasts isolated from spinach. The light-dependent decrease in (H⁺) in the chloroplast stroma counteracts carbon reduction and is offset at low light intensities by a large decrease in NADP and a significant increase in $\text{[}\text{ATP}\text{]}\text{[}\text{ADP}\text{]}$ ratios. Excess accumulation of NADPH and/or ATP permits 3-phosphogly cerate reduction to occur. With increasing light intensity, NADP levels and $\text{[}\text{ATP}\text{]}\text{[}\text{ADP}\text{]}$ ratios increased. High rates of photosynthesis were observed at high and at low $\text{[}\text{ATP}\text{]}\text{[}\text{ADP}\text{]}$ ratios. Levels of dihydroxyacetone phosphate were dramatically increased in the light. In chloroplasts, this permitted conversion to ribulose bisphosphate which on carboxylation yields 3-phosphoglycerate. The light-dependent alkalization of the chloroplast stroma is known to be responsible for phosphogly cerate retention in the chloroplasts. A high chloroplast ratio of phosphogly cerate to dihydroxyacetone phosphate aids carbon reduction. Measured ratios of dihydroxyacetone phosphate to phosphogly cerate were averages between low chloroplast ratios and high cytosolic ratios. They were far higher, even under low-intensity illumination, than dark ratios. Since cytosolic NADH levels are known to increase much less in the light than cytosolic dihydroxyacetone phosphate levels, the large increase in the ratio of didydroxyacetone phosphate to phosphogly cerate must considerably increase cytosolic phosphorylation potentials even at very low light intensities. It is proposed that this increase is communicated to the mitochondrial adenylate system, and inhibits dark respiratory activity, giving rise to the Kok effect. The extent of stroma alkalization, the efficiency of metabolite shuttles across the chloroplast envelope, and rates of cytosolic ATP consumption are proposed to be factors determining whether and to what extent the Kok effect can be observed. Light activation of chloroplast enzymes was slow at low and fast at high light intensities. This contrasts to low NADP levels at low and usually higher levels at high light intensities. Maximum enzyme activation was observed far below light saturation of photosynthesis, and light activation of enzymes was often less pronounced at very high than at intermediate light intensities.     

Regulation of nitrogenase activity in aerobes by MG2+ availability: An hypothesis

Nitrogenase functions at or near its maximum capacity $\text{in}\text{vivo}$, despite a reported energy charge in the cell that should severely inhibit the enzyme. Deenergizing cellular membranes, which is postulated to release magnesium in mitochondria, has been reported to produce rapid inhibition of nitrogenase activity while giving only small changes in energy charge and $\text{NAD}{}^{\mathrm{+}}\text{NADH}$ ratio. It is proposed that the level of magnesium available for complexation by the potent inhibitor ADP is the rate controlling variable for nitrogenase activity.     

Two-dimensional separation of chloroplast membrane proteins by isoelectri focusing and electrophoresis in sodium dodecyl sulphate

Proteins of chloroplast subfragments enriched in Photosystem I and Photosystem II electron flow activity have been analyzed by two-dimensional polyacrylamide gel electrophoresis. In the first dimension, polyacrylamide gel isoelectric focusing (pH 5–7) was used in the presence of Triton X-100, followed at right angle by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate. Characteristic fingerprints were obtained for the Photosystem I and II fractions and a correlation between the major proteins separated by isoelectric focusing and the major polypeptides separated by undimensional SDS electrophoresis was established. Two dominant spots of 68 000 and 60 000 daltons appeared in the two-dimensional patterns of Photosystem I fractions $\text{p}\text{I}$ values about 5.6; two spots with molecular weights of 33 000 and 23 000 were characteristics for Photosystem II fractions $\text{p}\text{I}$ values about 5.3 and 6.3). Photosystem I fractions were furthermore characteristics by a series of spots in the 44 000–33 000 range $\text{p}\text{I}$ values from about 5.9 to 6.8). The two-dimensional system revealed that (a) several SDS-polypeptides have multiple forms differing in charge only, (b) some proteins separated by isoelectric focusing are resolved in the second dimensional into polypeptides of different size. The two-dimensional method combining Triton X-100 isoelectric focusing' and SDS electrophoresis provides a higher degree of resolution than either of the unidimensional methods thus allowing a detailed analysis of chloroplast membrane proteins.     

Identification of a chloroplast membrane polypeptide associated with the oxidizing side of photosystem II by the use of select low-fluorescent mutants of Scenedesmus

Comparison of photochemical activities and variable fluorescence yield characteristics of whole cells and isolated chloroplast particles of low-fluorescent, photosystem II mutants of $\text{Scenedesmus}$$\text{obliquus}$ to those of the wild-type showed that several strains were affected primarily on the oxidizing side of photosystem II. In strains LF-1, LF-3, and LF-5 analysis of the manganese content of isolated chloroplast membranes showed a predominant shift in the $\text{Mn}\text{400}$ Chl from the wild-type value (4.3) to values near 1.5; this difference was also associated with a near total loss of cytochrome b-559 (high potential). Examination of chloroplast membrane polypeptides by gel electrophoresis revealed a decrease only in the mobility of one band in all three mutants; the apparent molecular weight was shifted from 34 kilodalton in the wild-type to 36 kilodalton in the mutants. Evidence is presented suggesting that the 34 kilodalton polypeptide of the wild-type is probably associated with the manganese requiring portion of the water-splitting apparatus of photosystem II.     

An analysis of proton fluxes coupled to electron transport and ATP synthesis in chloroplast thylakoids

Electron transport, phosphorylation and internal proton concentration were measured in illuminated spinach chloroplast thylakoid membranes under a number of conditions. Regardless of the procedure used to vary these parameters, the data fit a simple chemiosmotic model. Protons from Photosystem II did not appear to be utilized differently from those derived from Photosystem I. The maximal phosphorylation efficiency ($\text{P}\text{e}{}_2$) for photophosphorylation in washed thylakoids under oxidizing conditions is likely to be $\text{4}\text{3}$. This value is consistent with a proton-to-electron-pair ratio of 4 for electron flow through both photosystems and a proton-to-ATP ratio of 3 for the chloroplast proton-ATPase.     

Regulation of nitrogenase activity in soybean nodules by ATP and energy charge

Nonphosphorylating condition under anaerobiosis stopped nitrogenase activity in nodules of soybean ($\text{Glycine}$$\text{max}$ var. Chippewa) in less than three minutes and aeration quickly activated the enzyme. This stop-and-go treatment can be repeatedly applied on excised nodules, and a concomitant low-and-high ATP and energy charge (EC) was observed. After 2 minutes under anaerobiosis, nodule ATP and EC were decreased, respectively, to 20 and 40% of the control. These decreases were not as great with longer anaerobic treatments, and there was no change in the content of total adenosine phosphates. Oxygen enrichment (40%) stimulated the activity of nitrogenase by 2.5 fold in four minutes with a concomitant increase of ATP and EC by 40% and 14%, respectively, and an exhaustion of AMP. Longer treatments of oxygen enrichment lessened the initial effects. These findings indicate that ATP and energy charge probably regulate the activity of nitrogenase $\text{in}$$\text{vivo}$ and an active adenylate kinase must be operating in the nodules to maintain an energy supply for the basal metabolism and for the nitrogenase under temporary stressed conditions.     

Light scattering,chlorophyll fluorescence and state of the adenylate system in illuminated spinach leaves

Scattering of green light and chlorophyll fluorescence by spinach leaves kept in a stream of air or nitrogen were compared with leaf adenylate levels during illumination with blue, red or far-red light. Energy charge and ATP-ADP ratios exhibited considerable variability in different leaves both in the dark and in the light. Variability is explained by different possible states of the reaction oxidizing triose phosphate or reducing 3-phosphoglycerate. Except when oxygen levels were low, there was an inverse relationship between light scattering and chlorophyll fluorescence during illumination with blue or red light. When CO₂ was added to a stream of CO₂-free air, chlorophyll fluorescence increased, sometimes after a transient decrease, and both light scattering and leaf $\text{ATP}\text{ADP}$ ratios decreased. Similar observations were made when air was replaced by nitrogen under blue or high-intensity red light. Under these conditions, over-reduction caused inhibition of electron transport and phosphorylation in chloroplasts. However, when air was replaced by nitrogen during illumination with low-intensity red light or far-red light, light scattering increased instead of decreasing. Under these light conditions, $\text{ATP}\text{ADP}$ ratios were maintained in the light. They decreased drastically only after darkening. Although $\text{ATP}\text{ADP}$ ratios responded faster than light scattering or the slow secondary decline of chlorophyll fluorescence due to illumination, it appeared that in the steady state, light scattering and chlorophyll fluorescence are useful indicators of the phosphorylation state of the leaf adenylate system at least under aerobic conditions, when chloroplast and extrachloroplast adenylate systems can effectively communicate.     

Coordinate control of intermediary metabolism in rat liver by the insulin/glucagon ratio during starvation and after glucose refeeding. Regulatory significance of long-chain acyl-CoA and cyclic AMP.

The levels of serum insulin, glucagon, and free fatty acids (FFA) and the tissue concentrations of hepatic cyclic AMP, long-chain acyl-CoA (LCA), adenine nucleotides, inorganic phosphate, the intermediates of the Embden-Meyerhof pathway, the citric acid cycle (including acetyl-CoA and free CoA), and the cytoplasmic and mitochondrial redox couples were determined in the rat 12, 24, and 48 h after food withdrawal and 5, 10, 20, 40, 60, and 120 min after the refeeding of glucose. Using the measured metabolite contents in the liver, the alterations in the concentration of malate, oxaloacetate, citrate, and α-ketoglutarate and the changes in the energy state of the adenine nucleotide system and the redox state of the NAD system were attributed to the cytoplasmic and mitochondrial compartments by applying established calculation methods. Glucose refeeding provoked significant alterations in all parameters investigated. These changes occurred within minutes, reversing the hormone and metabolite pattern which had developed within 24 h in response to food withdrawal. Particularly, glucose refeeding resulted in a drastic increase in the insulin/glucagon ratio. Simultaneously, the level of serum FFA and the concentration of LCA in the liver declined. The latter alteration was accompanied by an increase in the cytoplasmic and a decrease in the mitochondrial $\text{ATP}\text{ADP}\text{×}\text{P}$ ratios. Moreover, the redox state of the cytoplasmic NAD system was shifted toward the oxidized state. When the appropriate data were plotted against each other, highly significant correlations were obtained (i) between the insulin/glucagon ratio and the serum FFA concentration, (ii) between the serum FFA concentration and the concentration of hepatic LCA, (iii) between the hepatic LCA concentration and the cytoplasmic energy state, and (iv) between the cytoplasmic energy state and the redox state of the cytoplasmic NAD system. These findings are interpreted to support the hypothesis derived from experiments carried out in vitro that the insulin/glucagon ratio via the FFA-dependent concentration of hepatic LCA might affect the translocation of adenine nucleotides between the cytoplasmic and the mitochondrial compartment, thereby regulating the cytoplasmic energy state and the redox state of the cytoplasmic NAD system, consequently. Glucose refeeding provoked rapid coordinate changes in the concentration of the intermediates of both the citric acid cycle and the Embden-Meyerhof chain, indicating the altered substrate flow through these pathways. Those metabolites, known to modulate the activity of key regulatory enzymes in vitro, were analyzed with respect to their suggested regulatory function. As to the established shift from pyruvate carboxylation to pyruvate decarboxylation after glucose refeeding, the data revealed that the decrease in pyruvate carboxylase activity can be attributed to the decrease in the intramitochondrial $\text{ATP}\text{ADP}$ ratio and the simultaneous fall in acetyl-CoA concentration, while the coordinate increase in pyruvate dehydrogenase activity can be ascribed to the decline in the concentration of LCA and, consequently, in the ratios of $\text{ATP}\text{ADP}$, $\text{NADH}\text{NAD}$, and acetyl-$\text{CoA}\text{CoA}$ within the mitochondria. As for the citric acid cycle, increased citrate synthesis from acetyl-CoA and oxaloacetate was supported by the rapid drop in the concentration of the established inhibitor of citrate synthesis, LCA. In contrast, the concentration of succinyl-CoA, an inhibitor of the enzyme in vitro, remained practically constant, questioning its regulatory function under the present experimental conditions. In addition to the activation of citrate synthase, the coordinate activation of isocitrate dehydrogenase was indicated by the LCA-mediated decline in both the mitochondrial $\text{ATP}\text{ADP}$ and the $\text{NADH}\text{NAD}$ ratios. Glucose refeeding immediately reduced urea excretion to basal values. This alteration was preceded by a drastic fall in the tissue concentration of cyclic AMP, supporting the physiological role of the nucleotide in the control of hepatic gluconeogenesis. In contrast, the observed changes in the concentration of the effectory acting metabolites (ATP, AMP, fructose 1,6-diphosphate, citrate, and alanine) were incompatible with the suggested function of these intermediates in switching over the substrate flow through the Embden-Meyerhof pathway from gluconeogenesis to glycolysis. The results are discussed in reference to the known rapid stimulation of fatty acid biosynthesis in the liver and to the transfer of reducing equivalents by the different shuttles of the inner mitochondrial membrane. In summary, it can be concluded that the insulin/glucagon ratio in a moment-to-moment fashion controls the glucose balance across the liver by regulating hepatic intermediary metabolism via the concentration of both LCA and cyclic AMP.     

Carbon monoxide and hydroxymercuribenzoate sensitivity of a fatty acid (omega-2) hydroxylase from Bacillus megaterium.

DNA-free minicells of $\text{Escherichia coli}$ will not allow growth of phage T-7, nor is RNA synthesis stimulated by phage infection. Thus, these miniature cells seem not to contain $\text{in vivo}$ RNA polymerase activity. However, DNA-dependent RNA polymerase activity can be unmasked in extracts with poly(dA-T) and Mn²⁺. This activity may represent a cytoplasmic pool of inactive RNA polymerase in normal cells.     

Effect of glucose on AMP-catabolite release during fatty acid perfusion in normal and ischemic rat hearts

Isolated rat hearts were perfused retrogradely with a modified, oxygenated Tyrode solution containing 0.5 mM palmitate (complexed to albumin in a molar ratio of 6:1) with or without 11mM glucose. Fatty acid perfusion induced a decrease in contractile behaviour which was partly counteracted by glucose. The energy charge $\text{{([}\text{ATP}\text{] +}\text{1}\text{2}\text{[}\text{ADP}\text{])/([}\text{ATP}\text{] + [}\text{ADP}\text{] + [}\text{AMP}\text{]}}$ of the tissue was not altered although a significant drop was observed in creatine phosphate/ATP ratio in the absence of glucose. The release of AMP-catabolites, adenosine, inosine and hypoxanthine, occurring during fatty acid perfusion was reduced by glucose. In the absence of glucose fatty acids still induce lactate release indicating an enhanced glycogenolysis. In ischemic hearts the fatty acid-induced decrease in mechanical performance was significantly more severe when glucose was absent, while the glucose protection could also be observed in the energy charge of the ischemic tissue and the release of AMP-catabolites in the coronary effluent. The results suggest that loss of adenosine, inosine and hypoxanthine might contribute to the detrimental actions of a high fatty acid/albumin ratio upon the myocardium and confirms the protective action of glucose.     

Euglena gracilis chloroplast ribosomes: improved isolation procedure and comparison of elongation factor specificity with prokaryotic and eukaryotic ribosomes

An improved method for the isolation of Euglena chloroplast ribosomes is described which presents a number of advantages over past procedures. First, ribosomes are prepared from whole cell extracts, thus bypassing the need to isolate intact chloroplasts and resulting in a 10-fold improvement in yield. Second, the inclusion of 40 mm Mg²⁺ in the preparation buffers, while stabilizing the chloroplast ribosomes, precipitates and, thereby, virtually eliminates the cytoplasmic 89 S ribosomes. Third, greater than 95% of the chloroplast ribosomes sediment at 68 S rather than as the damaged 53 S particle frequently generated in other preparation procedures. Fourth, even with a high-salt wash to remove endogenous factors, the chloroplast ribosomes still sediment at 68 S and are just as active in in vitro protein synthesis as are E. coli ribosomes. These ribosomes have been tested for activity with elongation factors from prokaryotes, eukaryotes, and the chloroplast itself, and the results have been compared to those obtained with E. coli and wheat germ ribosomes. The data may be summarized as follows: (a) Chloroplast ribosomes use E. coli$\text{EF}\text{-}\text{Tu}\text{Ts}$ and EF-G with the same efficiency as do E. coli ribosomes in protein synthesis, (b) E. coli and chloroplast ribosomes can use Euglena chloroplast EF-G to catalyze translocation, but wheat germ ribosomes cannot, (c) Wheat germ EF-1_H and EF-2 are highly active in polymerization with wheat germ ribosomes, but ribosomes from neither E. coli nor the chloroplast are able to recognize these factors, (d) All three types of ribosomes accept Phe-tRNA from E. coli EF-Tu although to differing degrees. However, neither chloroplast nor E. coli ribosomes recognize wheat germ EF-1_H for the binding of Phe-tRNA.     

ATP and adenylate energy charge during phosphate-mediated control of antibiotic synthesis.

The addition of inorganic phosphate or guanosine-5′-monophosphate to a phosphate-limited mycelial system of $\text{Streptomyces griseus}$ inhibited candicidin production. Accompanying the inhibition was a rapid increase in intracellular ATP concentration. Adenylate energy charge increased only slightly indicating that ATP is a more likely intracellular effector than energy charge in mediating phosphate control of antibiotic biosynthesis.