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41.
Paul Nieuwenhuysen Julius Clauwaert 《Journal of biochemical and biophysical methods》1981,5(5):279-286
It is impossible to measure the diffusion coefficient of macromolecules directly and accurately by quasi—elastic light scattering, when aggregates cannot be eliminated from the solutions to be investigated. Nevertheless, a simple method can be applied to overcome this problem in many cases. Aggregates are separated from the monomeric macromolecules by rate-zonal sedimentation in a CsCl density gradient in a transparent centrifugation tube; the monomers are then located by laser light scattering intensity measurements; photon correlation spectroscopy of the scattered light finally yields their diffusion coefficient. The viscosity of aqueous CsCl solutions at different temperatures and concentrations allows a good separation by centrifugation and a low uncertainty in the reduction of the measured diffusion coefficient to standard conditions.The application of the method to eukaryotic large ribosomal subunits is described as an example. 相似文献
42.
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 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, ratios were maintained in the light. They decreased drastically only after darkening. Although 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. 相似文献
43.
Noemi Luknar-Gabor Ursula Fenger Christoph Wagener Heinz Breuer 《Biochemical and biophysical research communications》1982,109(4):1270-1275
A variant of the carcinoembryonic antigen (CEA) with lower molecular weight than a CEA reference preparation has been separated from CEA. Using a polyclonal, spleen absorbed anti-CEA antiserum, the variant crossreacts with reference CEA in immunodiffusion. The CEA-activity of the variant has been demonstrated using an enzyme-immunoassay with monoclonal CEA specific antibodies. There is sufficient immunological evidence that this variant is a distinct antigen different from the crossreactive antigens described so far. The reactivity of the polyclonal anti-CEA antiserum with the CEA variant was abolished by absorption against the immobilized variant. 相似文献
44.
Yamazaki Y Hosono K Matsuda H Minami N Asai M Nakanishi H 《Biotechnology and bioengineering》1991,38(10):1218-1222
Twelve species of optically active metallocene derivatives having a 4-nitrophenyl group were prepared with horse liver alcohol dehydrogenase- or lipase-catalyzed optical resolution as the key step. The second harmonic generation (SHG) efficiently of these products was measured by the power method using the fundamental light from the Nd:YAG laser. (-)-1- (4-Nitrophenylthio)ethylferrocene (9), (+)-1-(4-nitrophenylthio)ethylruthenocene (16), (+)-1-(4-nitrophenylthio)ethylosmocene (19), (+) -1-(5-nitro-2-pyridylthio)ethylruthenocene (21), and (+) -1-[(4-nitrophenylhydrazono)methyl] -2-methylferrocene (12) showed SHG signals. The highest SHG efficiency was found with (+) -16, being 27 times more intense than the commonly used urea standard. 相似文献
45.
Shih-fang Fan M. M. Dewey B. Gaylinn B. Chu 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1992,162(6):508-512
Summary In dynamic light scattering, measurements of the intensity-intensity time correlation function from a suspension of rod-like particles of length L could reveal dynamical information related to translational and internal motions of those particles. For a suspension of thick filaments isolated from the myosin-regulated, striated muscles of Limulus at KL>1 (where K is the scattering vector), the average characteristic linewidth (
) increased with the addition of Ca2+ or with the depletion of ATP. The increase in the
with the addition of Ca2+ could be due to the presence of energy-requiring, high-frequency motions of the crossbridges activated by Ca2+. The increase in
which occurred with the depletion of ATP was assumed to be mainly due to the thermal motions of the crossbridges after they had moved radially away from the filament backbone. The percentage increase in
following the addition of Ca2+ was found to be seasonal, i.e., values of
obtained from thick filaments isolated between the middle of June and the middle of September were smaller than those obtained during the rest of the year. The effect of temperature on the percentage increase in
was also different. The increase showed a maximum at about 35°C during the summer and at about 25°C at other times. However, the percentage increase in
developed under ATP-depleted conditions showed no temperature-related maximum. The number of bound Ca2+ per myosin molecule was 1 during the summer and 2 at other times.Abbreviations DLS
dynamic light scattering
- L
length
- K
scattering vector
- SDS-PAGE
sodium dodecyl sulfate polyacrylamide gel electrophoresis
-
average characteristic line width
Deceased 相似文献
46.
Four chiral analogues of the surfactant Aerosol-OT (AOT) have been synthesized and characterized. All of them form reverse micelles in apolar solvents in the w0 range 0–30 (w0 = [water]/[tenside]). Reverse micellar solutions have been investigated by UV absorption and circular dichroism spectroscopies with the aim of clarifying whether the formation of the macromolecular micellar structure induces the appearance of new chromophoric bands or perturbs the existing ones. Methanolic solutions of the surfactants, in which no micellar aggregates are formed, were taken as references. One of the products 1(S),1′(S)-dimethylbisheptylsulphosuccinate sodium salt (MH-AOT) was capable of forming reverse micelles of relatively high water content (w0 up to 40) and this process was accompanied by a specific increase in the intensity of the circular dichroism band associated with the ester absorbance of the molecule. As no concomitant changes were seen in the UV absorbance spectrum, it was concluded that this observation reflected conformational events occurring within the surfactant rather than chromophoric perturbation. These results are qualitatively similar to those found recently for lecithin reverse micelles which, however, form gels at sufficiently high water contents. The chiroptical properties of these supramolecular aggregates are compared with those of covalent macromolecular systems such as polypeptides. 相似文献
47.
The β-carbon of the acyl group of β-hydroxy-β-methylglutarylhydroxyabscisic acid was shown to possess R-configuration by HPLC analysis of the reduced product. 相似文献
48.
Summary We have extended earlier experiments on the radiolysis of DL-tryptophan using32P-radiation to longer reaction times, observing complete destruction of the tryptophan by secondary, non-radiolytic processes. We have also undertaken the irradiation of DL-leucine with32P's at -196°, achieving radiolyses to the extents of ca. 20–30%, but observing no concomittant asymmetric bias. The implications of these observations are discussed with regard to the Vester-Ulbricht mechanism for the origin of optical activity. 相似文献
49.
GASTON DUCET 《Physiologia plantarum》1980,50(3):241-250
Oxidation of succinate by potato tuber mitochondria has been investigated from aerobiosis to complete anuerobiosis. Difference spectra of the various steps were recorded by a rapid scan spectrometer delivering averaged spectra every 3 s in the range 380 to 630 mm. The transitions between state 3 and 4 resulted in large redox changes, essentially for the b cytochromes, and in significant changes in the spectral baseline (light scattering). At anaerobiosis the cytochromes c, c1 and a were reduced while cytochrome a, remained oxidized. – Addition of uncouplers in aerobiosis induced oxidation of the b cytochromes, and when anaerobiosis occurred cytochromes c, c1a and a3 were reduced simultaneously. When uncouplers were added in anaerobiosis a partial oxidation of the b cytochromes and the reduction of cytochrome a3 were observed. These results are interpreted as the building up of a membrane potential, maximal in state 4 and stable after anaerobiosis. The cytochromes buried in the membrane equilibrate with the membrane potential, and their redox states are sensitive to the changes. Variations of membrane potential also induce changes in the light scattering by the mitochondrial membrane. 相似文献
50.
H. Notbohm 《Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression》1982,696(2):223-225
A DNA-protein complex isolated from Thermoplasma acidophilum has been examined using low-angle X-ray scattering measurements. In agreement with the results of electron-microscopic studies a diamter of 5.5 nm is deduced. Finally, a simplified model of the DNA-protein particles is discussed postulating a kinked DNA. 相似文献