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101.
102.
W D McCubbin G E Willick C M Kay 《Biochemical and biophysical research communications》1973,50(3):926-933
The fluorescent analog of adenosine triphosphate (ATP)1 1,N6-ethenoadenosine triphosphate, (εATP), has been utilized as a substitute for ATP in the myosin and heavy meromyosin ATPase systems. For myosin, the analog εATP replaced ATP with a somewhat larger Km (2.6 × 10?4 mole ??1 for εATP as opposed to 8.8 × 10?5 mole ??1 for ATP), indicating that the apparent affinity of the enzyme for εATP is less than for ATP. Perhaps of more interest, further comparison yielded a Vmax for εATP about two and one half times the value for ATP (20 μmole PO4 sec?1 g protein?1 as opposed to 8.1 μmole sec?1 g protein?1). Results for the HMM-εATPase system were similar, yielding a Km value of 1.47 × 10?4 mole ??1 and a Vmax of 54.2 μmole PO4 sec?1 g protein?1, as opposed to corresponding Km and Vmax values of 1.23 × 10?4 mole ??1 and 20.4 μmole PO4 sec?1 g protein?1, respectively for the HMM-ATP interaction. The pH dependence of εATPase for both systems was comparable to ATP, suggesting a similarity in the mechanism of hydrolysis of the two nucleotides. Activation of εATPase by Ca2+ in the presence of 0.5 M KCl was comparable to ATPase for both systems, but inhibition by Mg2+ seemed to be more effective for εATPase. These results indicate that εATP is an excellent substitute for ATP in the myosin and heavy meromyosin systems and because of its insertion into the active site of these muscle proteins, it promises to be a very useful probe for conformation studies at this level. 相似文献
103.
Acidic amino acid transport in Neurospora crassa mycelia 总被引:2,自引:0,他引:2
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107.
Properties of the transport systems for amino acids in Pseudomonas aeruginosa were investigated. Exogenous (14)C-labeled amino acids were shown to equilibrate with the internal native amino acid pool prior to incorporation into protein. When added at low external concentrations, the majority of the amino acids examined entered the protein of the cell unaltered. The rates of amino acid transport, established at low concentrations with 18 commonly occurring amino acids, varied as much as 40-fold. The transport process became saturated at high external amino acid concentrations, was temperature-sensitive, and was inhibited by sodium azide and iodoacetamide. Intracellular to extracellular amino acid ratios of 100- to 300-fold were maintained during exponential growth of the population in a glucose minimal medium. When the medium became depleted of glucose, neither extracellular nor intracellular amino acids could be detected. 相似文献
108.
The accumulation and behavior of various amino acids in the pool of Pseudomonas aeruginosa (ATCC 9027) were investigated. Patterns of pool formation and maintenance varied with different amino acids tested and were dependent, to a considerable extent, upon the ability of the organism to catabolize the particular amino acid. The establishment of steady-state amino acid pool levels depended upon the activity of the amino acid permease involved and upon the rate of protein synthesis. The presence of a relatively large specific amino acid pool did not affect the formation of a pool of a structurally different amino acid, and a preformed steady-state pool was not displaced by structurally unrelated amino acids. Steady-state amino acid pools decreased rapidly in the presence of inhibitors of energy metabolism and at 0 C. Steady-state internal amino acid pools were found to be in equilibrium with the corresponding external amino acid, present at low levels. A multiplicity of proline pools was demonstrated. 相似文献
109.
Comparison of the Action of Colicins E1 and K on Escherichia coli with the Effects of Abortive Infection by Virulent Bacteriophages 总被引:11,自引:1,他引:10
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Kay L. Fields 《Journal of bacteriology》1969,97(1):78-82
Abortive infection of certain strains of Escherichia coli or Shigella dysenteriae with phages of the T-even group or with phage T5 resembles the action of colicin E1 or K on sensitive bacteria, especially in the effects on biosynthetic processes. Tests on transport systems and on adenosine triphosphate levels suggest, however, that different mechanisms are involved in the two cases. Abortive infection appears to cause damage to the permeability barrier of the cell, whereas the colicins interfere more directly with the energy metabolism of the bacteria. 相似文献
110.
Colicins E1 and K inhibited a whole series of energy-dependent reactions in Escherichia coli cells, including motility, biosynthesis of nucleic acids, proteins and polysaccharides, and the conversion of ornithine to citrulline. Respiration was only partially affected, and substrates such as glucose continued to be catabolized through the normal pathways, albeit with reduced CO(2) production. The soluble products of aerobic glucose catabolism by colicin-treated cells were analyzed. Pyruvate replaced acetate as the major excreted product, and the following intermediates of glycolysis were excreted in significant amounts: glucose-6-phosphate, fructose-1,6-diphosphate, dihydroxyacetone phosphate, and 3-phosphoglycerate. Anaerobically growing cells manifested a somewhat enhanced tolerance to the colicins. This protection by anaerobiosis appeared to depend on the exclusion of oxygen more than on the extent of fermentative catabolism versus catabolism of the respiratory type. These results are interpreted in terms of possible functions of colicin in lowering the adenosine triphosphate (ATP) content of the cells and in terms of the role of lowered ATP levels in inhibiting many of the energy-requiring reactions. 相似文献