• 자연과학논문집
• /
• 제8권2호
• /
• pp.35-41
• /
• 1996

MCAT1 의 기질과 결합하는 장소를 조사하기 위해 세포막내에 존재하는 부분의 극성 아미노산을 반대 극성의 아미노산으로 치환시킨 돌연변이체를 만들어 arginine 통과능력, gp70 결합능, efflux, Xenopus oocyte 를 이용한 단백질 발현 능력등을 조사하였다. 다섯 개의 돌연변이체중 D403K 이 가장 흥미로운 성질을 나타냈는데, 그것은 정상적인 gp70 결합능력과 낮은 arginine 통과능력을 나타냈다는 것으로 정상적인 단백질이 세포막에서 발현되고 있으나 arginine 통과능력이 감소했다는 사실을 나타내는 것이다. K211E 를 제외한 모든 돌연변이체가 감소된 arginine efflux 와 감소된 Vmax 값을 나타내었다. 이들 결과로부터 MCAT1 의 403 번째 아미노산인 glutamic acid 가 세포막내에서 기질과 상호작용하는 장소라는 가능성을 시사하였다.

• 한국물환경학회지
• /
• 제22권3호
• /
• pp.566-571
• /
• 2006

A sequencing batch biofilm reactor (SBBR) operated with a cycle of anaerobic - aerobic - anoxic - aerobic has been evaluated for the nutrient removal characteristics. The sponge-like moving media was filled to about 10% of reactor volume. The sewage was the major substrate while external synthetic carbon substrate was added to the anoxic stage to enhance the nitrogen removal. The operational results indicated that maximum T-N and T-P removal efficiencies were 97% and 94%, respectively were achieved, while COD removal of 92%. The observations of significant nitrogen removal in the first aerobic stage indicated that nitrogen removal behaviour in this SBBR was different to conventional SBR. Although the reasons for aerobic nitrogen removal has speculated to either simultaneous nitrification and denitrification or anoxic denitrification inside of the media, further researches are required to confirm the observation. The specific oxygen uptake rate (SOUR) test with biofilm and suspended growth sludge indicated that biofilm in SBBR played a major role to remove substrates.

• Archives of Pharmacal Research
• /
• 제25권6호
• /
• pp.984-988
• /
• 2002

The issue of whether or not the presence NOx (NO and oxidized metabolites) in the hepatocytes at pathological levels affects the functional activity of transport systems within the sinusoidal membrane was investigated. For this purpose, the effect of the pretreatment of isolated hepatocytes with sodium nitroprusside (SNP), a spontaneous NO donor, on the sinusoidal uptake of tributylmethylammonium (TBuMA) and triethylmethyl ammonium (TEMA), representative substrates of the organic cation transporter (OCT), and taurocholate, a representative substrate of the $Na^+$/taurocholate cotransporting polypeptide (NTCP), was measured. The uptake of TBuMA and TEMA was not affected by the pretreatment, as demonstrated by the nearly identical kinetic parameters for the uptake ($i.e., V_{max}, K_{m} and CL_{linear}$). The uptake of mannitol into hepatocytes was not affected, demonstrating that the membrane integrity remained constant, irregardless of the SNP prutreatment. On the contrary, the uptake of taurocholate was significantly inhibited by the pretreatment, resulting in a significant decrease in V_{max}$, thus providing a clear demonstration that NOx preferentially affects the function of NTCP rather than OCT on the sinusoidal membrane. A direct interaction between NOx and NTCP or a decrease in $Na^+/K^+$ ATPase activity as the result of SNP pretreatment might be responsible for this selective effect of NOx.

• Environmental Engineering Research
• /
• 제11권3호
• /
• pp.156-163
• /
• 2006

In this study, substrate removal characteristics were analyzed to reduce the cost of external carbon dosage at Sudokwon Landfill Site Management Corporation in Korea by utilizing oxygen uptake rate (OUR) and nitrate uptake rate (NUR) tests. To estimate and evaluate the substrate removal characteristics obtained by the batch tests, the lab-scale MLE process was operated. By-products of J Co. (sugar manufactory) and S Co. (fine chemical industry) were selected as the concerned carbon sources through a comparison of carbon and nitrogen contents. MeOH was tested as a control experiment. Until the steady state, the fraction of $RBDCOD_{OUR}$ concentration to COD concentration of J Co., S Co. by-products and MeOH increased and reached levels of 98%, 82%, and 100%, respectively. During the 20th operating day, the fraction of $RBDCOD_{NUR}$ concentration to COD concentration was 95%, 81%, and 83%, respectively. These fractions of $RBDCOD_{NUR}$ concentration to $RBDCOD_{OUR}$ concentration increased according to acclimation periods and reached levels of 99%, 97%, and 81%, respectively, on the 20th day. The results obtained from the lab-scale MLE process operation using the concerned carbon sources as external carbon were similar to that observed by OUR and NUR tests.

• Archives of Pharmacal Research
• /
• 제18권1호
• /
• pp.12-17
• /
• 1995

It is essential to clone the preptide transporter in order to obtain better understanding of its molecular structure, regulation, and substrate specificity. Characteristics of an endogenous peptide transporter in oocytes were studied along with expression of an exogenous protor/peptide cotransporter from rabbit intestine. And further efforts toward cloning the transporter were performed. The presence of an endogenous peptide transporter was detected in Xenopus laevis oocytes by measuring the uptake of $0.25/{mu}M(10{\;}{\mu}Ci/ml)[^3H]$-glycylsarcosine (Gly-Sar) at pH 5.5 with or without inhibitors. Yptake of Gly-Sae in oocytes was significantly inhibited by $25{\mu}M$ glycine nd sarcosine. This result suggests that a selective transporter is involved in the endogneous uptake of dipeptides. Collagenase treatment of oocytes used to strip oocytes from ovarian follicles did not affect the Gly-Sar uptake. Changing pH from 5.5 to 7.5 did not affect the Gly-Sae uptake significantly, suggesting no depedence of the endogenous transporter on a transmembrane proton gradient. An exogenous $H^+/pep-tide$ contransported was expressed after microinjection of polyadenylated messenger ribonucleic acid $[poly(A)^+ -mRNA]$ obtained from rabbit small intestine. The Gly-Sar uptake in mRNA-injected oocytes was 9 times thigher than that in water-injected oocyltes. Thus, frog occytes can be utilized fro expression cloning of the genes encoding intestinal $H^+$peptide contransporters. Size fractionation of mRNA was successfully obtained using this technique.

• 대한의생명과학회지
• /
• 제15권1호
• /
• pp.55-60
• /
• 2009

Insect cells such as Spodoptera frugiperda Clone 9 (Sf9) cells are widely chosen as the host for heterologous expression of a mammalian sugar transport protein using the baculovirus expression system. Characterization of the expressed protein is expected to include assay of its function, including its ability to transport sugars and to bind inhibitory ligands such as cytochalasin B. It is therefore very important first to establish the transport characteristics and other properties of the endogenous sugar transport proteins of the host insect cells. However, very little is known of the transport characteristics of Sf9 cells, although their ability to grow on TC-100 medium strongly suggested the presence of endogenous glucose transport system. In order to investigate the substrate and inhibitor recognition properties of the Sf9 cell transporter, the ability of pentoses to inhibit 2-deoxy-D-glucose (2dGlc) transport was investigated by measuring inhibition constants $(K_i)$. To determine the time period over which of sugar into the Sf cells was linear, the uptake of 2dGlc 0.1mM extracellular concentration was measured over periods ranging from 30 seconds to 30 minutes. The uptake was linear for at least 2 minutes at the concentration, implying that uptake made over a 1 minute time course would reflect initial rates of the sugar uptake. The data have also revealed the existence of a saturable transport system for pentose uptake by the insect cells. The transport was inhibited by D-xylose and D-ribose, although not as effective as hexoses. However, L-xylose had a little effect on 2dGlc transport in the Sf9 cells, indicating that the transport is stereoselective. Unlike the human erythrocyte-type glucose transport system, D-ribose had a somewhat greater apparent affinity for the Sf9 cell transporter than D-xylose. It is therefore concluded that Sf9 cells contain an endogenous sugar transport activity that in some aspects resembled the human erythrocyte-type counterpart, although the Sf9 and human transport systems do differ in their affinity for cytochalasin B.

• 생명과학회지
• /
• 제32권10호
• /
• pp.762-770
• /
• 2022

제 2형 당뇨병은 고혈당을 특징으로 하는 만성 대사성 질환으로 인슐린 민감성및 저항성을 개선하여 세포속으로 포도당 흡수를 촉진시킴으로서 완화될 수 있다. 본 연구는 triterpenoid 화합물인 betulinic acid가 3T3-L1 지방세포에서 인슐린 신호전달체계를 개선하여 포도당 흡수를 촉진시키는지를 조사하고 그 작용기전을 규명하였다. Betulinic acid는 3T3-L1 지방세포에서 농도의존적으로 포도당 흡수를 유의하게 증가시켰으며, 이는 PM-GLUT4의 발현 증가와 관련이 있음을 관찰하였다. Betulinic acid는 인슐린 신호전달 경로에서 PI3K의 활성화 및 IRS-1tyr, Akt의 인산화를 대조군에 비해 유의하게 증가시켰다. 또한 AMPK의 활성화를 나타내는 pAMPK와 AMPK 하위인자인 pACC의 수준을 유의하게 증가시켰다. Betulinic acid에 의한 PI3K 및 AMPK 경로의 활성화를 증명하기 위해, PI3K 억제제(Wortmannin)와 AMPK의 억제제(Compound C)를 사용하여 이들 처리에 의한 포도당 흡수능과 PM-GLUT4의 발현을 측정한 결과 이들의 발현이 유의하게 저해되었다. 본 연구에서 betulinic acid는 3T3-L1 지방세포에서 PI3K 및 AMPK 경로의 활성화를 통해 세포막으로 포도당 수송체인 GLUT4 전위를 촉진시키고 포도당 흡수를 증가시킬 수 있음을 나타내었다. 이러한 결과는 betulinic acid가 인슐린 민감성을 증진시키고 고혈당을 완화하는데 도움이 될 수 있음을 시사한다.

• The Korean Journal of Physiology and Pharmacology
• /
• 제6권1호
• /
• pp.21-26
• /
• 2002

Effects of cadmium (Cd) intoxication on renal endosomal accumulation of organic cations $(OC^+)$ were studied in rats using $^{14}C-tetraethylammnium$ (TEA) as a substrate. Cd intoxication was induced by s.c. injections of 2 mg Cd/kg/day for $2{\sim}3$ weeks. Renal cortical endosomes were isolated and the endosomal acidification (acridine orange fluorescence change) and TEA uptake (Millipore filtration technique) were assessed. The TEA uptake was an uphill transport mediated by $H^+/OC^+$ antiporter driven by the pH gradient established by $H^+-ATPase.$ In endosomes of Cd-intoxicated rats, the ATP-dependent TEA uptake was markedly attenuated due to inhibition of endosomal acidification as well as $H^+/TEA$ antiport. In kinetic analysis of $H^+/TEA$ antiport, Vmax was reduced and Km was increased in the Cd group. Inhibition of $H^+/TEA$ antiport was also observed in normal endosomes directly exposed to free Cd (but not Cd-metallothionein complex, CdMt) in vitro. These data suggest that during chronic Cd exposure, free Cd ions liberated by lysosomal degradation of CdMt in proximal tubule cells may impair the endosomal accumulation of $OC^+$ by directly inhibiting the $H^+/OC^+$ antiporter activity and indirectly by reducing the intravesicular acidification, the driving force for $H^+/OC^+$ exchange.

• BMB Reports
• /
• 제32권5호
• /
• pp.429-435
• /
• 1999

Unlike the mammalian glucose transporter GLUT1, little is known about the nature of the endogenous sugar transporter(s) in insect cells. In order to establish the transport characteristics and other properties of the sugar transport proteins of Sf9 cells, a series of kinetic analyses was performed. A saturable transport system for hexose uptake has been revealed in the insect cells. The apparent affinity of this transport system(s) for 2-deoxy-D-glucose was relatively high, the $K_m$ for uptake being <0.5 mM. To further investigate the substrate and inhibitor recognition properties of the insect cell transporter, the ability of other sugars or drugs to inhibit 2-deoxy-D-glucose transport was examined by measuring inhibition constants ($K_j$). Transport was inhibited by D-mannose, D-glucose, and D-fructose. However, the apparent affinity of the C-4 epimer, D-galactose, for the Spodoptera transporter was relatively low, implying that the hydroxyl group at the C-4 position may play a role in the strong binding of glucose and mannose to the transporter. The results also showed that transport was stereoselective, being inhibited by D-glucose but not by L-glucose. It is therefore concluded that insect cells contain an endogenous glucose transport activity that in several aspects resembles the human erythrocyte glucose transporter. However, the mammalian and insect transporters were different in some of their kinetic properties, namely, their affinities for fructose and for cytochalasin B.

• Journal of Ecology and Environment
• /
• 제38권2호
• /
• pp.123-131
• /
• 2015

During several visits to the Cebu City landfill in the Philippines, plants were observed growing within the area, including on top of the garbage piles. Studying the response of these plants is important in assessing which can be used in remediating metal contaminated soils. This study aimed to determine whether the plants in the Cebu City landfill excluded or accumulated cadmium (Cd) and chromium (Cr) in the plant tissues. The floristic composition of the landfill was analyzed prior to the sample collection. The samples were acid-digested before the desired elements were measured using atomic absorption spectrophotometry (AAS). The Cd and Cr concentrations in the plant root-zone soil were also measured using AAS. The results indicated that the landfill substrate was generally acidic based on the results of the pH measurement. Of the 32 plant species sampled, Cyperus odoratus showed potential for Cd uptake and internal transfer; Cenchrus echinatus, Vernonia cinerea and Terminalia catappa for Cr uptake, and Cynodon dactylon for Cr internal transfer. The plants in the landfill differed in their response towards the heavy metals. To confirm the behavior of C. odoratus towards Cd, and C. echinatus, C. dactylon, V. cinerea, and T. catappa towards Cr, controlled experiments are recommended, as the plant samples analyzed were collected from the field.