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81.
Danuta Dus Maciej Ugorski Wojciech Gorczyca Czeslaw Radzikowski 《Glycoconjugate journal》1989,6(3):355-365
125I-Wheat germ agglutinin (WGA) binding parameters of human urothelial cell lines of different grades of transformation (TGrll and TGrlll) were compared. The values of association constant (Ka) and the number of binding sites/cell for HCV29 (TGrll) cell line were about 3×106M–1 and over 4×107, respectively. Two TGrlll cell lines, HCV29T and Hu549 revealed lower values for Ka, and considerably higher numbers of binding sites/cell (about 3×108 and 2×108, respectively). Binding of125I-WGA to total cellular proteins resolved by SDS-PAGE and transferred to nitrocellulose showed multiple diffused bands in the range of 58–180 kDa. Some of these bands were characteristic for TGrll cells (124 kDa) or TGrlll cells (135 and 148 kDa).Abbreviations TGr
transformation grade
- WGA
wheat germ agglutinin
- sWGA
succinylated wheat germ agglutinin
- GlcNAc
N-acetyl-d-glucosamine
- BSA
bovine serum albumin
- SDS-PAGE
sodium dodecyl sulphate polyacrylamide gel electrophoresis 相似文献
82.
Scott Brian Lew James Clandinin M. T. Cinader B. 《Cellular and molecular neurobiology》1989,9(1):105-113
1. SJL/J mice were maintained on semipurified diets which differed in the ratio of polyunsaturated/saturated fatty acid content (P/S). Exposure was from conception and was maintained for periods ranging from 6 to 34 weeks. 2. Neural cell cultures were prepared from dorsal root ganglia (DRG). After 6 and 20 days of culture, neuronal electric membrane properties were determined quantitatively by intracellular recording. 3. A number of significant differences were observed for the two dietary conditions. DRG from mice on the low-P/s diet had an increase in the rate of fall of both phases of repolarization which, in conjunction with the reduced action potential overshoot, led to a reduced action potential duration. This shift to shorter-duration action potentials was accompanied by a shift to more monophasic falling phases. The low-P/S neurons also exhibited a decreased afterhyperpolarization, decreased specific membrane resistance, and decreased membrane electrical time constant compared to high-P/S neurons. 4. It was concluded that the P/S ratio in the diet can have a significant effect on the electric properties of neurons. The high-P/S neurons tended to have action potentials with biphasic repolarizations and longer durations. In contrast, the low-P/S neurons tended to have action potentials with monophasic repolarizations and shorter durations. Moreover, the known ionic dependence of these two types of action potentials suggested that the low-P/S diet resulted in action potentials with a more exclusive Na dependence, while the high-P/S diet resulted in action potentials with both Na and Ca dependence. 相似文献
83.
Shi Liu Ron Jacob David Piwnica-Worms Melvyn Lieberman 《Molecular and cellular biochemistry》1989,89(2):147-150
We have recently reported the presence of an electroneutral (Na + K + 2 Cl) cotransport mechanism that is bumetanide-sensitive and maintains Cli above its electrochemical equilibrium in cultured chick heart cells. In steady state, (Na + K + 2 Cl) cotransport is inwardly directed and so contributes to the Na influx that must be counterbalanced by the activity of the Na/K pump to maintain Nai homeostasis. We now show that manipulating (Na + K + 2 Cl) cotransport by restoring Clo to a Cl-free solution indirectly influences Na/K pump activity because the bumetanide-sensitive recovery of a
infNa
supi
to its control level and the accompanying hyperpolarization could be blocked by 10–4M ouabain. In another protocol, when the Na/K pump was reactivated by restoring Ko (from 0.5 mM to 5.4 mM) and removing ouabain, the recovery of aNa was attenuated by 10–4M bumetanide. The relatively slow rate of ouabain dissociation coupled with the activation of Na influx by (Na + K + 2 Cl) cotransport clearly establishes the interaction of these transport mechanisms in regulating Nai. Although (Na + K + 2 Cl) cotransport is electroneutral, secondary consequences of its activity can indirectly affect the electrophysiological properties of cardiac cells. 相似文献
84.
Vijittra Leardkamolkarn Louis W. Heck Dale R. Abrahamson Ph.D. 《Cell and tissue research》1989,257(3):587-596
Summary Laminin biosynthesis and basement membrane assembly in anterior pituitary glands of gonadectomized rats were studied by immuno-electron microscopy and radioimmunoassay. Three weeks after gonadectomy, rats received intravenous injections of sheep anti-laminin IgG conjugated to horseradish peroxidase, and glands were fixed and processed for microscopy 1 h later. Peroxidase reaction product uniformly labeled all perivascular and glandular epithelial basement membranes. In addition, reaction product was also found in abnormally multi-layered basement membranes seen especially beneath gonadotrophs, and unusual basement membrane-like structures projecting between gonadotrophs were also labeled. Pituitary sections from gonadectomized rats labeled with pre-embedding immunoperoxidase and post-embedding immungold techniques also localized intracellular laminin within biosynthetic organelles and light body vesicles of gonadotrophs. Neither abnormal basement membrane structures nor intracellular laminin were detected in pituitaries of nongonadectomized, control rats. Radioimmunoassays of pituitary homogenates showed nearly twice as much soluble laminin ( 15 ng/gland) in gonadectomized rats than in controls ( 8 ng/gland), which paralleled gland growth, but serum laminin concentrations did not differ ( 10 ng/ml in both groups). When anterior pituitary glands of gonadectomized rats that received injections of anti-laminin IgG-HRP were fixed 5 days after injection, lengths of unlabeled basement membrane were distributed between labeled lengths. This indicated that new basement membrane was spliced into old by a process similar to that seen in normal development. Supplementation of gonadectomized rats with testosterone, however, arrested laminin biosynthesis and basement membrane assembly and reversed glandular hypertrophy. These results indicate that, in an absence of sex hormone feedback, renewed synthesis of basement membrane components occurs in the anterior pituitary and is probably necessary to support the additional growth and differentiation of gonadotrophs and other pituitary cells. 相似文献
85.
Rolf Mentlein Cornelia Buchholz Prof. Dr. Brigitte Krisch 《Cell and tissue research》1989,258(2):309-317
Summary The synthetic peptides somatostatin (SRIF) and growth hormone-releasing hormone (GRH) were coupled directly to colloidal gold of different particle sizes. Both conjugates were biologically active in displacing the corresponding radiolabeled hormones from high affinity binding sites in pituitary membranes. Release of growth hormone (GH) from cultured anterior pituitary cells was modulated by both conjugates alone or in combination. Ultrastructural studies were performed with cells incubated at 4° C (2 h) and 37° C (2 min-2 h) with one of the labeled peptides or their combination. Somatotropes were identified by immunostaining with anti-rGH followed by protein A-ferritin, thus obtaining a triple labeling. Both hormone conjugates were internalized in different vesicles in the beginning but accumulated during longer incubation times in the same compartment. The secretory vesicles and the nucleus were not labeled by any hormone conjugate. In contrast to SRIF-gold, the uptake of GRH-gold conjugate decreased with longer incubation times. This effect could be neutralized by simulatenous incubation of the somatotropes with both regulating hormones. Hence, whereas the binding and internalization of SRIF by somatotropes do not seem to be influenced by GRH, the corresponding processes for GRH are stimulated by the presence of SRIF. 相似文献
86.
H+/sugar symport in the obligatory aerobic yeastRhodotorula glutinis was analyzed under conditions where the plasma membrane was selectively depolarized by the lipophilic cation tetraphenylphosphonium (TPP+). Control experiments showed that this treatment did not impair the transmembrane pH, the cell energy charge, and the function of plasma membrane H+-ATPase. The kinetic data were fitted to elementary functions derived from a model constructed on the basis of some simplifying premises for ordered (either C + H+ + S or C + S + H+) and random reaction mechanisms. In addition, the comparison of the kinetic parameters in fully energized and depolarized cells provided information about the free carrier charge. It was concluded that the binding sequence of formation of the ternary carrier/H+/substrate complex follows a random mechanism and that the carrier bears a negative charge. 相似文献
87.
Recent experiments show that membrane ATPases are capable of absorbing free energy from an applied oscillating electric field and converting it to chemical bond energy of ATP or chemical potential energy of concentration gradients. Presumably these enzymes would also respond to endogenous transmembrane electric fields of similar intensity and waveform. A mechanism is proposed in which energy coupling is achieved via Coulombic interaction of an electric field and the conformational equilibria of an ATPase. Analysis indicates that only an oscillating or fluctuating electric field can be used by an enzyme to drive a chemical reaction away from equilibrium.In vivo, the stationary transmembrane potential of a cell must be modulated to become locally oscillatory if it is to derive energy and signal transduction processes. 相似文献
88.
E. C. Foulkes 《Biological trace element research》1989,21(1):195-200
Review of the available evidence on the mechanism of cellular Cd uptake in the rat jejunum supports the concept that this
process consists of nonspecific binding to anionic sites on the membrane, followed by a temperature-dependent and rate-limiting
internalization step. Because temperature-sensitive transmembrane movement of Cd can be demonstrated also in isolated brush-border
vesicles and in erythrocyte ghosts, it is not likely to result from pinocytosis but may be related directly to membrane fluidity.
There is no need to assume the existence of saturable Cd carriers, or competition of Cd with essential polyvalent cations
for their specific transport systems. Uptake of Cd by tubular epithelium in the kidney of the intact rabbit appears to resemble
that described for the jejunum, with the internalization step limiting the rate of uptake. 相似文献
89.
Denis Roy Marco Lagimonire Marie-Jose Hardy Jean-Franois Bourassa Pierre Mourot 《Journal of biotechnology》1989,10(3-4):227-240
Factors affecting the viability and infectivity of an ectomycorrhizal fungus during moderate concentration by cross-flow filtration were determined. Mycelial suspensions were concentrated with three commercial membrane filters (Prostak Millipore Co., M14 Tech-Sep Co. and Ceraflo Norton Co.) under aseptic conditions. Medium components may reduce the filtration rate due to their low solubility. An antifoam agent did not reduce the average flux rates as much as did the malt extract. Clear unobstructed channels (I.D. 6mm) of the tubular modules (Tech-Sep) gave the best results both in terms of performance (filtration rate) and cell viability. Shear stresses caused by pumping and flow through narrow retentate channels were probably responsible for lowering viability and infectivity. There was no linear relationship between permeate fluxes and cell concentration. There is an optimum pore size both in terms of performance (filtration rate) and cell viability. Physical blockage of large pores by hyphae could explain lower permeate flux rates than those obtained with lower pore sizes membranes. 相似文献
90.
C. Bevan R. K. H. Kinne R. E. Shetlar E. Kinne-Saffran 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1989,159(3):339-347
Summary A membrane fraction, rich in brush border membranes, was prepared from renal proximal tubules of the spiny dogfish,Squalus acanthias, and the sodium-proton exchange mechanism in these membrane vesicles was investigated by both a rapid filtration technique and the fluorescence quenching of acridine organe.22Na+ uptake was stimulated by an outwardly directed H+ gradient, and was inhibited by amiloride at a single inhibitory site with an apparentK
i
of approximately 1.7×10–5
M. In the presence of an H
i
+
>H
o
+
gradient, the
of the Na+/H+ exchanger were 9.7±0.8 mM and 48.0±12.0 nmol·mg protein–1·min–1, respectively. The uptake of Na+ was electroneutral in the presence of a H+ gradient, indicating a stoichiometry of 1. In the fluorescence studies, quenching of acridine orange occurred in the presence of an outwardly directed Na+ gradient which was inhibited by amiloride. Thus, an electroneutral Na+/H+ exchanger with properties similar to those found in the mammalian kidney is also present in the spiny dogfish and may contribute to the urinary acidification of this marine animal. 相似文献