• Bulletin of the Korean Chemical Society
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• v.26 no.4
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• pp.675-678
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• 2005

• Bulletin of the Korean Chemical Society
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• v.32 no.8
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• pp.2791-2794
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• 2011

• Bulletin of the Korean Chemical Society
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• v.25 no.5
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• pp.721-725
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• 2004

Acetohydroxy acid synthase (AHAS) is one of several enzymes that require thiamine diphosphate and a divalent cation as essential cofactors. The active site contains several conserved ionizable groups and all of these appear to be important as judged by the fact that mutation diminishes or abolishes catalytic activity. Recently, we have shown [Yoon, M.-Y., Hwang, J.-H., Choi, M.-K., Baek, D.-K., Kim, J., Kim, Y.-T., Choi, J.-D. FEBS Letters 555 (2003), 185-191] that the activity is pH-dependent due to changes in $V_{max}$ and V/$K_m$. Data were consistent with a mechanism in which substrate was selectively catalyzed by the enzyme with an unprotonated base having a pK 6.48, and a protonated group having a pK of 8.25 for catalysis. Here, we have in detail studied the pH dependence of the kinetic parameters of the cofactors (ThDP, FAD, $Mg^{2+}$) in order to obtain information about the chemical mechanism in the active site. The $V_{max}$ of kinetic parameters for all cofactors was pH-dependent on the basic side. The pK of ThDP, FAD and $Mg^{2+}$ was 9.5, 9.3 and 10.1, respectively. The V/$K_m$ of kinetic parameters for all cofactors was pH-dependent on the acidic and on the basic side. The pK of ThDP, FAD and $Mg^{2+}$ was 6.2-6.4 on the acidic side and 9.0-9.1 on the basic side. The well-conserved histidine mutant (H392) did not affect the pH-dependence of the kinetic parameters. The data are discussed in terms of the acid-base chemical mechanism.

• The Korean Journal of Physiology and Pharmacology
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• v.23 no.5
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• pp.381-392
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• 2019

Sperm function and male fertility are closely related to pH dependent $K^+$ current (KSper) in human sperm, which is most likely composed of Slo3 and its auxiliary subunit leucine-rich repeat-containing protein 52 (LRRC52). Onion peel extract (OPE) and its major active ingredient quercetin are widely used as fertility enhancers; however, the effect of OPE and quercetin on Slo3 has not been elucidated. The purpose of this study is to investigate the effect of quercetin on human Slo3 channels. Human Slo3 and LRRC52 were co-transfected into HEK293 cells and pharmacological properties were studied with the whole cell patch clamp technique. We successfully expressed and measured pH sensitive and calcium insensitive Slo3 currents in HEK293 cells. We found that OPE and its key ingredient quercetin inhibit Slo3 currents. Inhibition by quercetin is dose dependent and this degree of inhibition decreases with elevating internal alkalization and internal free calcium concentrations. Functional moieties in the quercetin polyphenolic ring govern the degree of inhibition of Slo3 by quercetin, and the composition of such functional moieties are sensitive to the pH of the medium. These results suggest that quercetin inhibits Slo3 in a pH and calcium dependent manner. Therefore, we surmise that quercetin induced depolarization in spermatozoa may enhance the voltage gated proton channel (Hv1), and activate non-selective cation channels of sperm (CatSper) dependent calcium influx to trigger sperm capacitation and acrosome reaction.

• Nuclear Engineering and Technology
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• v.9 no.2
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• pp.75-81
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• 1977

This experiment has been carried out to determine the pH dependent cation exchange capacity concerning the sorption phenomenon of long-lived radionuclides contained in low-level liquid radioactive waste on various clay minerals. The pH dependent cation exchange capacity determined by Sawhney's method are used to the analysis of sorption phenomenon. About 70 percent of the total cation exchange capacity is contributed by the pH dependent CEC due to the negative charge originated naturally in clays in case of clinoptilolite, vermiculite and sodalite. It is sugested in this test that the high neutral salt CEC, that is, highly charged clays would show good fixation yield. The removal of radionuclides at the pH range more than pH 9 is considered the hydroxide precipitation of metal ion rather than the cation exchange. The Na-clay prepared by the method of successive isomorphic substitution with electrolyte showed a considerable improvement in removal efficiency for the decontamination.

• Macromolecular Research
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• v.13 no.2
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• pp.147-151
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• 2005

The main objective of this study was to modulate pH-sensitive polymers (poly($\beta-amino esters$)) by controlling their molecular weight during their synthesis. These pH-sensitive and biodegradable polymers were synthesized by Michael-type step polymerization. 1,4-Butane diol diacrylate was used as the unsaturated carbonyl compound and piperazine as the nucleophilic compound. Various molecular weight polymers were obtained by varying the mol ratio of piperazine/1,4-butane diol diacrylate. The synthesized polymers were characterized by $^{1}H-NMR$ and their molecular weights were measured by gel permeation chromatography(GPC). The dependence of the molecular weight on the mol ratio was evaluated by the titration method. Also, the pH dependent turbidity of the polymers was determined by UV-Vis spectrophotometry. This pH dependent property of the polymers could be very useful for preparing drug carriers that are sensitive to pH.

• Proceedings of the Korean Society of Applied Pharmacology
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• 2001.11a
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• pp.88-88
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• 2001

The effects of pH and temperature on the degradation of prokidn in various buffered aqueous solutions(pH 1.32~9.66) and temperatures (35, 45 and 6$0^{\circ}C$ were investigated. The effect of ionic strength on the degradation of prokidin was also measured by varying ionic strength (0.0466~1.5) at pH 1.35 and 45$^{\circ}C$ The effect of metal ions on the degradation of prokidin at pH 7.35 and 3.98 was observed. The degradation of prokidin followed the pseudo- first- order kinetics. The degradation rate of prokidin showed pH-dependent and temperature-dependent patterns. Prokidin was very stable at the pH below 3.95, where half-lives at 35, 45 and 6$0^{\circ}C$were 294, 206 and 107 day, respectively. However, it degraded very rapidly at pH above 6.49; the half-lives at 35, 45 and 6$0^{\circ}C$were 60, 25 and 13 day, respectively. As ionic strength increased, the degradation rate of prokidin increased. Some metal ions increased the degradation rate in the rank order of Mn > Fe > Cu >Fe On the other hand. other metal ions such as Bi, Ba. Zn, Ni, Co did not show unfavorable effect.

• Parasites, Hosts and Diseases
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• v.27 no.3
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• pp.161-170
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• 1989

The proteases of Toxoplasma gcndii were purified partially and characterisrd for some biochemical properties including various chromatographic patterns, major catalytic classes, and conditions to promote the activity of these enzymes. When Toxoplasma extract was incubated with 3H-casein at various pH, peak hydrolysis of casein was observed at pH 6.0 and at pH 8.5. Proteasfs working at pH 6.0 and at pH 8.5 were purified partially by conventional methods of chromatographies of DE52 anion rxchange, Sephadex G-200 gel permeation, and hydroxylapatite chromatography. Partially purified enzymes were tested by site-specific inhibitors and promotorf. The protease working at pH 6.0 was inactivated by iodoacetamide with LDso of 10-5 M and promoted by dithiothreitol, while the protease working at pH 8.5 was inhibited by phenylmethylsulfonyl fluoride with LD50 of 10-5 M and was Promoted by ATP (excess ATP beyond 2 mM inhibited the activity reversely). The protease of pH 8.5 had the activity of ATPase which might exert the energy to its action. Therefore the former was referred to as a cysteinyl acid protease and the latter, ATP-dependent neutral serine protease.

• Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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• v.2 no.2
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• pp.135-143
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• 2004

An experimental study on the sorption of U(VI) onto a Korean granite was performed as a function of the geochemical parameters such as contact time, pH, ionic strength, and carbonate concentration using a batch procedure. The distribution coefficient,$K_d$, was about 1-200 mL/g depending on the experimental conditions. The sorption of U(VI) onto granite particles was greatly dependent upon the contact time, pH, and carbonate concentration, but insignificantly dependent on the ionic strength. It was noticed that the sorption of U(VI) onto granite particles was highly correlated with the uranium speciation in the solution, which was dependent on the pH and carbonate concentrations. It was deduced from the kinetic sorption experiment that a two-step first-order kinetic behavior could dominate the kinetic sorption of U(VI) onto granite particles. In the alkaline range of a pH above 7, U(VI) sorption was greatly decreased and this might be due to the formation of anionic U(VI)-carbonate aqueous complexes as predicted by the speciation calculations.

• The Korean Journal of Physiology and Pharmacology
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• v.21 no.2
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• pp.241-249
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• 2017

Plasma membrane hyperpolarization associated with activation of $Ca^{2+}$-activated $K^+$ channels plays an important role in sperm capacitation during fertilization. Although Slo3 (slowpoke homologue 3), together with the auxiliary ${\gamma}^2$-subunit, LRRC52 (leucine-rich-repeat-containing 52), is known to mediate the pH-sensitive, sperm-specific $K^+$ current KSper in mice, the molecular identity of this channel in human sperm remains controversial. In this study, we tested the classical $BK_{Ca}$ activators, NS1619 and LDD175, on human Slo3, heterologously expressed in HEK293 cells together with its functional interacting ${\gamma}^2$ subunit, hLRRC52. As previously reported, Slo3 $K^+$ current was unaffected by iberiotoxin or 4-aminopyridine, but was inhibited by ~50% by 20 mM TEA. Extracellular alkalinization potentiated hSlo3 $K^+$ current, and internal alkalinization and $Ca^{2+}$ elevation induced a leftward shift its activation voltage. NS1619, which acts intracellularly to modulate hSlo1 gating, attenuated hSlo3 $K^+$ currents, whereas LDD175 increased this current and induced membrane potential hyperpolarization. LDD175-induced potentiation was not associated with a change in the half-activation voltage at different intracellular pHs (pH 7.3 and pH 8.0) in the absence of intracellular $Ca^{2+}$. In contrast, elevation of intracellular $Ca^{2+}$ dramatically enhanced the LDD175-induced leftward shift in the half-activation potential of hSlo3. Therefore, the mechanism of action does not involve pH-dependent modulation of hSlo3 gating; instead, LDD175 may modulate $Ca^{2+}$-dependent activation of hSlo3. Thus, LDD175 potentially activates native KSper and may induce membrane hyperpolarization-associated hyperactivation in human sperm.