• Animal cells and systems
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• v.2 no.4
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• pp.539-543
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• 1998

Hrp1, of Schizosaccharomyces pombe, is a new member of the SW12/SNF2 protein family that contains a chromodomain and a DNA binding domain as well as ATPase/7 helicase domains. This configuration suggests that Hrp1 could be a homolog of mouse CHD1, which is thought to function in altering the chromatin structure to facilitate gene expression. To understand the enzymatic nature of Hrp1 we purified the 6-Histidine-tagged Hrp1 protein (6$\times$His-Hrp1) to homogeneity from a S. pombe Hrp1-overexpressing strain and hen examined its biochemical properties. We demonstrate that the purified 6$\times$His-Hrp1 protein exhibited a DNA-binding activity with a moderate preference to the (A＋T)-rich tract in double-stranded NA via a minor groove interaction. However, we failed to detect any intrinsic DNA helicase activity from the purified Hrp1 like other SW12/SNF2 proteins. These observations suggest that the DNA binding activities of Hrp1 may be involved in the remodeling of the chromatin structure with DNA-dependent ATPase. We propose that Hrp1 may function in heterochromatins as other proteins with a chromo- or ATPase/helicase domain and play an important role in the determination of chromatin architecture.

• Journal of Photoscience
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• v.4 no.2
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• pp.41-48
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• 1997

The plasma membrane and microsomes, isolated from the cells treated with hematoporphyrm derivative (HpD) for 1 and 24 h, accumulated the aggregated porphyrin. The quantity of aggregated porphyrin was same in the plasma membrane and microsomes after isolating them from cells treated with HpD for 1 h whereas the microsomes accumulated higher quantity of aggregated porphyrin when cells were treated with HpD for 24 h. Photodynamic action of aggregated porphyrin on plasma membrane and microsomes was investigated using lipid specific fluorescent probes: 1,6-diphenyl-1,3,5-hexatrine (DPH) and 1-(4-trimethylammonium), 6-diphenyl-1,3,5-hexatrine(TMA-DPH). The time dependent anisotropy of these probes in the membranes was measured and the decay of anisotropy was analyzed using wobbling in cone model. Upon irradiation both the plasma membrane and the microsomes showed an increase in the limiting anis~)tropy and order parameter and a decrease in the cone angle of the lipid probes. The increase in the limiting anisotropy was pronounced in membranes isolated from the cells treated with HpD for 24 h. Photoinduced change in the limiting anisotropy was dependent on the duration of incubation of cells with HpD before isolating the membranes. In both the membranes. the membrane core was affected more as compared to the outer leaflet. In addition to the structural changes, a decrease in Na$^+$-K$^+$-ATPase and NADPH cyt c reductase activity was also observed upon irradiation of HpD treated cells. Inhibition in NADPH cyt c reductase was more when cells were treated with HpD for 24 h, however, Na$^+$-K$^+$-ATPase activity did not depend on the duration of the treatment of cells with HpD before irradiation. Our results suggest that the extent of photoinduced perturbations in the membranes varies as a function of duration of the treatment of cells with HpD and the membrane core is more susceptible to the photodynamic action of aggregated porphyrin.

• Korean Journal of Clinical Laboratory Science
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• v.41 no.2
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• pp.83-92
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• 2009

This study was undertaken to examine the effect of oxidants on membrane transport function in renal epithelial cells. Hydrogen peroxide ($H_2O_2$) was used as a model oxidant and the membrane transport function was evaluated by measuring $Na^+$-dependent phosphate ($Na^+$-Pi) uptake in opossum kidney (OK) cells. $H_2O_2$ inhibited $Na^+$-Pi uptake in a dose-dependent manner. The oxidant also caused loss of cell viability in a dose-dependent fashion. However, the extent of inhibition of the uptake was larger than that in cell viability. $H_2O_2$ inhibited $Na^+$-dependent uptake without any effect on $Na^+$-independent uptake. $H_2O_2$-induced inhibition of $Na^+$-Pi uptake was prevented completely by catalase, dimethylthiourea, and deferoxamine, suggesting involvement of hydroxyl radical generated by an iron-dependent mechanism. In contrast, antioxidants Trolox, N,N'-diphenyl-p-phenylenediamine, and butylated hydroxyanisole did not affect the $H_2O_2$ inhibition. Kinetic analysis indicated that $H_2O_2$ decreased Vmax of $Na^+$-Pi uptake with no change in the Km value. Phosphonoformic acid binding assay did not show any difference between control and $H_2O_2$-treated cells. $H_2O_2$ also did not cause degradation of $Na^+$-Pi transporter protein. Reduction in $Na^+$-Pi uptake by $H_2O_2$ was associated with ATP depletion and direct inhibition of $Na^+$-$K^+$-ATPase activity. These results indicate that the effect of $H_2O_2$ on membrane transport function in OK cells is associated with reduction in functional $Na^+$-pump activity. In addition, the inhibitory effect of $H_2O_2$ was not associated with lipid peroxidation.

• Korean Journal of Microbiology
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• v.30 no.6
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• pp.528-532
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• 1992

The ATP-dependent protease. Clp P from Esehaichia coli has been increase the stahility with or without detergent as Triton X-100 and NP-40 in the Clp P. The C]p P proteolytic activity was remained to 0.1 M salt by $Na^{-1}$, $K^{+}$, $Li^{+}$ but was inhihited by $SO_4^{2}$. An active ATPase site in Clp A is required for A TP-dependent proteolysis by Clp protease as

• Journal of Microbiology
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• v.37 no.2
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• pp.59-65
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• 1999

RecA protein can promote strand assimilation, homologous pairing, and strand exchange. All these reactions require DNA-dependent ATP hydrolysis by recA protein, and the activities of recA protein are affected by the ionic environment. In this experiment, DNA-dependent ATPase activity showed different sensitivity to anionic species. ATP hydrolysis and strand exchange were relatively sensitive to salt in the reactions with NaCl, strongly inhibited at 100 mM NaCl. However, the inhibition by sodium acetate or sodium glutamate was not observed at 50∼100 mM concentration. Addition of sodium glutamate to the standard reaction condition increased the apparent efficiency of ATP hydrolysis during strand exchange. The condition including 50∼100 mM sodium-glutamate might be similar to the physiological condition.

• The Korean Journal of Physiology and Pharmacology
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• v.3 no.5
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• pp.529-538
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• 1999

This study was undertaken to examine the effect of ethanol on $Na^+ -dependent$ phosphate $(Na^+-P_i)$ uptake in opossum kidney (OK) cells, an established renal proximal tubular cell line. Ethanol inhibited ^Na^+-dependent$ component of phosphate uptake in a dose-dependent manner with $I_{50}$ of 8.4%, but it did not affect $Na^+-independent$ component. Similarly, ethanol inhibited $Na^+-dependent$ uptakes of glucose and amino acids (AIB, glycine, alanine, and leucine). Microsomal $Na^+-K^+-ATPase$ activity was not significantly altered when cells were treated with 8% ethanol. Kinetic analysis showed that ethanol increased $K_m$ without a change in $V_{max}$ of $Na^+-P_i$ uptake. Inhibitory effect of n-alcohols on $Na^+-P_i$ uptake was dependent on the length of the hydrocarbon chain, and it resulted from the binding of one molecule of alcohol, as indicated by the Hill coefficient (n) of 0.8-1.04. Catalase significantly prevented the inhibition, but superoxide dismutase and hydroxyl radical scavengers did not alter the ethanol effect. A potent antioxidant DPPD and iron chelators did not prevent the inhibition. Pyrazole, an inhibitor of alcohol dehydrogenase, did not attenuate ethanol-induced inhibition of $Na^+-P_i$ uptake, but it prevented ethanol-induced cell death. These results suggest that ethanol may inhibit $Na^+-P_i$ uptake through a direct action on the carrier protein, although the transport system is affected by alterations in the lipid environment of the membrane.

• The Korean Journal of Physiology
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• v.16 no.2
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• pp.177-186
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• 1982

The effect of ethacrynic acid (EA) on the renal secretion of PAH was examined in cat kidney. $C_{PAH}$ and $T_{PAH}$ were measured before and after infusion of EA $(0.5{\sim}50mg/kg)$ through the femoral vein. The following results were obtained: 1) In the dosage range of 0.5 to 25 mg/kg, EA increased the urine flow, and sodium and potassium excretion in dose-dependent manner, but the glomelular filtration rate was decreased as the dosage of EA was increased. 2) $C_{PAH}$ and $T_{PAH}$ were decreased by EA in the dosage range of 3 to 25 mg/kg and 1 to 50 mg/kg, respectively, in dose·dependent manner with the dosage to cause 50% inhibition of about 5 mg/kg. 3) With dosage of 0.5mg/kg, EA appeared to exert a great effect on diuretic response without the influence on $T_{PAH}$. At 10min after infusion of EA, a potent diuretic effect appeared, while $T_{PAH}$ did not show a significant change. These results suggest that the action mechanism of EA on tubular secretion of PAH may be different from that on natriuresis. 4) With dosage of 5 mg/kg, EA did not inhibit the Na-K-ATPase activity in microsomal fractions from both cortex and medulla. 5) The double reciprocal plot ($l/T_{PAH}$ versus $l/P_{PAH}$) suggested that EA inhibited the P AH secretion by a competitive pattern. However, probenecid, a prototypic inhibitor of the organic acid pump, had no influence on both the inhibitory effect of $T_{PAH}$ and the natriuretic effect by EA. These results suggest that in vivo EA altered tubular secretion of P AH through interactions with receptors that are not identical with the Na-K-ATPase.

• Korean Journal of Food Science and Technology
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• v.40 no.4
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• pp.424-429
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• 2008

To investigate the acid tolerance characteristics of the acid-resistant mutant, Leuconostoc paramesenteroides P-200, as a kimchi starter, this study examine proton permeability, ATPase activity, glycolysis activity, $Mg^{2+}$ release, and membrane fatty acid composition, and compared the data to that of its wild-type, L. paramesenteroides LP-W. In the proton permeability experiment, the LP-W and P-200 strains' average maximum half-time $(t_{1/2})$ values for pH equilibration through the cell membrane were approximately 5.7 and 9.3 min in 150mM KCl solution, and 4.2 and 8.3 min in 3% NaCl solution, respectively. Their values and pH levels for maximal specific ATPase activity showed that P-200 had greater activity than LPW. And the results of pH-dependent glycolysis activity showed that P-200 had greater activity than LP-W. Furthermore, after 2 hr at pH 4.0, LP-W and P-200 had percent magnesium release values of approximately 12% and 34%, respectively. A comparison of their membrane fatty acid compositions indicated that C18 and cyclo-C19 were the major different fatty acids between the two strains, and their contents of C18 and cyclo-C19 were 2.5% and not detected, respectively, in LP-W, and 6.4% and 11.4%, respectively, in P-200. These results indicate that the P-200 strain has significantly improved acid tolerance as compared to its wild type, LP-W.

• Journal of Ginseng Research
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• v.41 no.4
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• pp.556-565
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• 2017

Background: Recent studies have revealed that the properties Traditional Chinese Medicine is mostly associated with are substance and energy metabolism. Our study aimed to compare the effect of red ginseng (RG) (warm property) and ginseng leaves (GL; cold property) on the substance and energy metabolism of rats with hypothyroidism. Materials and methods: Rats were administered propylthiouracil intraperitoneally for 20 d to cause hypothyroidism. The reference group was orally administered Aconiti Lateralis Radix Praeparaia [FZ (Fuzi in Chinese)], while both the RG and GL groups were orally administrated crude drugs. The rectal, tail, toe, and axilla temperature of the rats were assayed every 3 d. Oxygen consumption, carbon dioxide production, heat production, and energy expenditure were measured via TSE phenoMaster/LabMaster animal monitoring system. Adenosine monophosphate-activated protein kinase, $Na^+-K^+$-ATPase, fumarase, pyruvic acid and cyclic adenosine monophosphate/cyclic guanosine monophosphate were determined. Results: The lower levels of triiodothyronine, tetraiodothyronine, and thyrotropin-releasing hormone and the higher level of thyroid stimulating hormone revealed the successful establishment of a hypothyroidism model. Oxygen consumption, carbon dioxide production, heat production, and energy expenditure in the FZ and RG groups were obviously increased. The activity of $Na^+-K^+$-ATPase and fumarase in the FZ and RG groups was significantly increased. The cyclic adenosine monophosphate/cyclic guanosine monophosphate level in the FZ and RG groups was increased, while the GL group showed the opposite. Conclusion: Our research provides a new way to explore the efficiency of Chinese medicine on the basis of the relationship between drug property and effects on substance and energy metabolism.

• Ocean and Polar Research
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• v.38 no.2
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• pp.103-113
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• 2016

This study aimed to examine the role of two aquaporin isoforms (AQP3 and AQP8) in response to the hyperosmotic challenge of transitioning from freshwater (FW) to seawater (SW) during parr and smoltification (smolt) using the rainbow trout, Oncorhynchus mykiss. We examined the changes in the expression of AQPs mRNAs in the gills and intestine of the parr and smolt stages of rainbow trout transferred from FW to SW using quantitative real-time PCR in an osmotically changing environment [FW, SW, and recombinant AQP3 (rAQP3) injection at two dosage rates]. Correspondingly, AQPs were greater during smoltification than during parr stages in the rainbow trout. Plasma osmolality and gill $Na^+/K^+$-ATPase activity increased when the fish were exposed to SW, but these parameters decreased when the fish were exposed to SW following treatment with rAQP3 during the transition to seawater. Our results suggest that AQPs play an important role in water absorbing mechanisms associated with multiple AQP isoforms in a hyperosmotic environment.