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91.
A new adsorption chromatography procedure for the purification of calmodulin from bovine brain was developed using polymeric adsorbent 3520. Calmodulin was first isolated by DEAE-Cellulose column chromatography and further purified to apparent homogeneity following elution with 50% ethanol from the adsorbent column. Polyacrylamide gel electrophoresis showed one band either in the presence of Ca2+ or EGTA. The polymeric adsorbent 3520 is a non-polar polymer lacking exchangeable groups. The selective adsorption of calmodulin is based on hydrophobic interaction within the matrix, and is Ca2+ independent. Neither high salt (0.5 M NaC1) nor EGTA (5 mM) was able to elute the CaM from the adsorption column whereas ethanol (50%) eluted it completely. This method is simple to use and it provides highly purified calmodulin with high yield. 相似文献
92.
93.
94.
Twelve- and sixteen-residue peptides have been designed to form tetrameric alpha-helical bundles. Both peptides are capable of folding into amphiphilic alpha-helices, with leucyl residues along one face and glutamyl and lysyl residues along the opposite face. Four such amphiphilic alpha-helices are capable of forming a noncovalently bonded tetramer. Neighboring helices run in antiparallel directions in the design, so that the complex has 222 symmetry. In the designed tetramer, the leucyl side chains interdigitate in the center in a hydrophobic interaction, and charged side chains are exposed to the solvent. The designed 12-mer (ALPHA-1) has been synthesized, and it forms helical aggregates in aqueous solution as judged by circular dichroic spectroscopy. It has also been crystallized and characterized by x-ray diffraction. The crystal symmetry is compatible with (but does not prove) the design. The design can be extended to a four-alpha-helical bundle formed from a single polypeptide by adding three peptide linkers. 相似文献
95.
The relationship between the activity of ATC oxygenase, CTC production and growth rate was investigated in a low-producing
strain ofStreptomyces aureofaciens, closely related to the wildtype strain, and in a higher-producing variant. Different growth rates were achieved by using
glucose, fructose and sucrose as carbon sources. Activity of the enzyme and CTC yield in both strains were inversely proportional
to the rate of sugar utilization but in the higherproducing variant sugar utilization was slower than in the low-producing
strain. The expression of ATC oxygenase was less sensitive to “catabolite repression” in the higherproduc ing strain. BT,
a stimulator of CTC production, markedly inhibited growth of the higher-producing variant in a medium with slowly utilized
sugars (fructose and sucrose) but had little effect on growth of the lowproducing strain. It also increased the level of ATC
oxygenase in both strains under all experimental conditions. It could be established that there was no obligatory relationship
between the increase of antibiotic synthesis and the increase of enzyme activity. 相似文献
96.
The inhibitory effect of trifluoroperazine (25-200 microM) on the sarcoplasmic reticulum calcium pump was studied in sarcoplasmic reticulum vesicles isolated from skeletal muscle. It was found that the lowest effective concentrations of trifluoroperazine (10 microM) displaces the Ca2+ dependence of sarcoplasmic reticulum ATPase to higher Ca2+ concentrations. Higher trifluoroperazine concentrations (100 microM) inhibit the enzyme even at saturating Ca2+. If trifluoroperazine is added to vesicles filled with calcium in the presence of ATP, inhibition of the catalytic cycle is accompanied by rapid release of accumulated calcium. ATPase inhibition and calcium release are produced by identical concentrations of trifluoroperazine and, most likely, by the same enzyme perturbation. These effects are related to partition of trifluoroperazine ino the sarcoplasmic reticulum membrane, and consequent alteration of the enzyme assembly within the membrane structure, and of the bilayer surface properties. The effect of trifluoroperazine was also studied on dissociated ('chemically skinned') cardiac cells undergoing phasic contractile activity which is totally dependent on calcium uptake and release by sarcoplasmic reticulum, and is not influenced by inhibitors of slow calcium channels. It was found that trifluoroperazine interferes with calcium transport by sarcoplasmic reticulum in situ, as well as with the role of sarcoplasmic reticulum in contractile activation. 相似文献
97.
Metabolic acidosis produces a phosphaturia which is independent of parathyroid hormone or dietary phosphorus intake. To study the underlying mechanism, inorganic phosphate (Pi) and glucose transport were studied in brush-border membrane vesicles prepared from the renal cortex of parathyroidectomized rats gavaged for three days with either 7.5 ml of 1.6% NaCl (control) or 1.5% NH4Cl (acidosis). At killing, blood pH and plasma bicarbonate were and , respectively, in control and () and () in acidotic rats. Serum Pi was similar in both groups, while 24 h urine Pi excretion was higher in the acidotic group (). Peak sodium-dependent uptake of Pi, measured after 1.5 min of incubation, was higher in controls than acidotic rats ( vs. protein, ), whereas peak glucose uptake at 1.5 min was not significantly different between the groups. Equilibrium values for Pi and glucose uptake were similar in the two groups. for Pi uptake in the control and acidotic animals were not different, 0.036 and 0.040 mM, respectively. By contrast, was higher in controls than in the acidotic group, 3.13 vs. 1.15 nmol/mg protein per 15 s. These results suggest that metabolic acidosis directly inhibits Pi uptake by the brush border of the proximal tubule by decreasing the availability of Pi carriers of the renal brush-border membrane. 相似文献
98.
Three photosynthetic enzymes were characterised in extractsfrom leaves and aerial roots of Aranda Christine 130.The enzymes from both tissues were similar in activity and kineticproperties. Grana-containing chloroplasts were found in rootcells of Vanda suauis. Thus components crucial to photosynthesisare present in aerial roots of these leafy orchids. (Received March 22, 1983; Accepted July 7, 1983) 相似文献
99.
D G Davis T R Lindstrom N H Mock J J Baldassare S Charache R T Jones C Ho 《Journal of molecular biology》1971,60(1):101-111
100.