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981.
In previous studies on the quaternary structure of Na+,K+-ATPase, cupric-phenanthroline complex (CP) has been used for the cross-linking of the enzyme subunits. Here we show that the same products obtained in the presence of CP (α,α-dimer, α,β-dimer, and products of higher molecular weight) are also obtained when the enzyme is exposed to Cu2+ without o-phenanthroline. The α,β-dimer (but not the α,α-dimer) is dissociated in the presence of EDTA; indicating that this product is not the result of the covalent cross-linking of the subunits through a disulfide bond. The nature of the α,α-dimer remains to be determined. The findings suggest that the results of “cross-linking” experiments with CP should be interpreted with caution until the products are more clearly identified. 相似文献
982.
A patient with cervical carcinoma was found to have selective IgA deficiency. The intact cell-mediated immunity, normal levels of IgG and IgM, and the absence of serum and salivary IgA established the diagnosis. Contrary to those of normal persons, salivary IgM was elevated and salivary IgA was not detectable in this patient. The patient had no signs attributable to IgA deficiency, but she always had dryness of the mouth. The association between cervical carcinoma and selective IgA deficiency was discussed. 相似文献
983.
984.
985.
986.
Summary The effects of various agents on active sodium transport were studied in the toad bladder in terms of the equivalent circuit comprising an active conductanceK
a, an electromotive forceE
Na, and a parallel passive conductanceK
p. For agents which affectK
a, but notE
Na orK
p, the inverse slope of the plot of total conductance against short-circuit currentI
0 evaluatesE
Na, and the intercept representsK
p. Studies employing 5×10–7
m amiloride to depressK
a indicate a changingE
Na, invalidating the use of the slope technique with this agent. An alternative suitable technique employs 10–5
m amiloride, which reducesI
0 reversibly to near zero without effect onK
p. Despite curvilinearity of the -I0 plot under these conditions,K
p may therefore be estimated fairly precisely from the residual conductance. It then becomes possible to follow the dynamic behavior ofK
a andE
Na (in the absence of 10–5
m amiloride) by frequent measurements of andI
0, utilizing the relationshipsK
a=K-K
p, andK
Na=I
O/(K-K
p). 2-deoxy-d-glucose (7.5×10–3
m) depressedK
a without affectingE
Na. Amiloride (5×10–7
m) depressedK
a and enhancedE
Na. Vasopressin (100 mU/ml) enhancedK
a markedly and depressedE
Na slightly. Ouabain (10–4
m) depressed bothK
a andE
Na. All of the above effects were noted promptly;K
p was unaffected. The electromotive force of Na transportE
Na appears not to be a pure energetic parameter, but to reflect kinetic factors as well, in accordance with thermodynamic considerations. 相似文献
987.
J P Leroux J C Marchand R Hong Tuan Ha P Cartier 《European journal of biochemistry》1975,58(2):367-373
Viable human polymorphonuclear leukocytes isolated from peripheral blood were incubated for 1 h at 37 degrees C with variable concentrations of insulin in a saline medium buffered at pH 7.4. The hormone increased glucose consumption by about 40% without influencing the permeability of the membranes to glucose, whose uptake followed a passive diffusion process. The measurement of intermediates localized activation of glycolysis by insulin, down to 0.36 nM, at the phosphofructokinase step. However, the spectrophotometric measurement showed no activation of phosphofructokinase after preincubation with insulin of either intact granulocytes or crude or ultracentrifuged homogenates. The level of cyclic AMP, which is known to activate phosphofructokinase, was not modified by insulin; cyclic GMP did not activate the enzyme in the granulocyte extracts: neither of the two nucleotides can therefore be considered as a direct messenger of the action of insulin on phosphofructokinase. An important fraction of the extra glucose consumed under the influence of insulin was recovered as neither glycogen nor lactate, nor was it oxidized in the Krebs cycle. It might be assumed to have been converted into glycerolipids. However, insulin produced no detectable accumulation of triglycerides and activated neither the pentose phosphate pathway nor oxidative decarboxylation of pyruvate. The fate of the extra glucose consumed under the influence of insulin therefore remains questionable. 相似文献
988.
The insoluble component of stratum corneum of rat epidermis yields two major bands after extraction with 8 M urea-mercaptoethanol-dithiothreitol. The ratio of these two bands is about 1:1 in terms of protein stain intensity and S-[14C]carboxymethyl label. Both polypeptides were purified to homogeneity by DE-52-cellulose, sodium dodecyl sulfate hydroxylapatite C column chromatography, and preparative DodSO4-polyacrylamide gel electrophoresis. The heavier polypeptide contains 30% alpha helix and the lighter contains 27% alpha helix as determined by circular dichroism studies. Both are sensitive to Pronase and resistant to trypsin, collagenase, and elastase. The lighter chain is stable to pepsin but the heavier can be partially degraded to a smaller polypeptide with a molecular weight similar to that of light chain. Amino acid analysis shows that the light chain contains 12 more tyrosine residues than does the heavy chain, suggesting that the light chain is not generated from the heavy chain. However, the two chains may have a common peptide region. Antiserum prepared against the heavier polypeptide can be completely absorbed by purified lighter polypeptide and vice versa indicating that both chains have some common antigenic determinants. Antibody against either chain can cross-react with the stratum corneum and keratohyalin granules in the epidermis of newborn rat as indicated by fluorescent microscopic observation. Similarly, this antibody also cross-reacts with the cell surface or the contents of spinous and granular cells, and very weakly with basal cells, indicating that the two proteins may be present in the lower strata as well as the stratum corneum. 相似文献
989.
Data are presented which prove that 3-O-methylfluorescein phosphate is a substrate for the K+-dependent phosphatase that is associated with Na+,K+-ATPase. Conditions for the continuous fluorimetric assay of 3-O-methylfluorescein phosphatase are described. Enzyme preparations from three different tissues with widely different specific activities exhibit similar Km values for 3-O-methylfluorescein phosphate. Correlation between Na+,K+-ATPase activity and K+-dependent 3-O-methylfluorescein phosphatase activity is demonstrated in several partially purified enzyme preparations and crude tissue fractions. When the K+-dependent 3-O-methylfluorescein phosphatase of a crude rat-brain homogenate is assayed, the activity is a linear function of the amount of homogenate added to the assay mixture. The equivalent of 10 μg of brain tissue may be assayed under the conditions used. The potential value of this highly sensitive fluorimetric method for the assay of enzyme in small samples of various tissues is suggested. 相似文献
990.
The characterization of hemoglobin Wood (beta97(FG4) His replaced by Leu), a high oxygen affinity hemoglobin with reduced Hill constant is described. The amino acid substitution occurs at the alpha1beta2 interface, in the same position as in hemoglobin Malm? (beta97(FG4) His replaced by Gln) and in an homologous position when compared with hemoglobins Chesapeake (alpha92(FG4) Arg replaced by Leu) and J. Capetown (alpha92(fg4) arg replaced by Gln). 相似文献