• Title/Summary/Keyword: Amino Acid Transport

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Determination of Analytical Approach for Ambient PM2.5 Free Amino Acids using LC-MSMS (LC-MSMS를 이용한 대기 중 PM2.5 유리아미노산 분석 방법 연구)

  • Bae, Min-Suk;Park, Da-Jeong;Lee, Kwon-Ho;Cho, Seung-Sik;Lee, Kwang-Yul;Park, Kihong
    • Journal of Korean Society for Atmospheric Environment
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    • v.33 no.1
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    • pp.54-63
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    • 2017
  • Atmospheric nitrogen containing organic compounds(e.g. amino acids) has attracted considerable attention from the viewpoint of the oceanic biogeochemical cycle of nitrogen as well as the long range transfer. However, only a few measurements of organic nitrogen compounds have been conducted due to analytical difficulties. In this study, total of nine amino acids such as Glutamic acid, Histidine, Arginine, Tyrosine, Cystine, Valine, Methionine, Phenylalanine, Lysine have been analytically determined by Liquid Chromatography - Mass Spectrometry Mass Spectrometry (LC-MSMS). As results, Fragmentor Voltage (FV), Precursor Ion, Collision Energy, Product Ion related to individual amino acid compounds are shown. Based on the operational conditions, Lysine, Glutamine Acid, Tyrosine were analyzed during the China Oriented Smog Period. High concentrations of Lysine, Glutamine Acid, and Tyrosine are discussed with organic carbon (OC), elemental carbon (EC), and water soluble ions. The results can provide to understand the sources with aging process related to amino acids influenced by the long-range transport from the Yellow Sea area.

Sodium Dependent Taurine Transport into the Choroid Plexus, the Blood-Cerebrospinal Fluid Barrier

  • Chung, Suk-Jae;Ramanathan, Vikram;Brett, Claire M.;Giacomini, Kathleen M.
    • Journal of Pharmaceutical Investigation
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    • v.25 no.3
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    • pp.7-20
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    • 1995
  • Taurine, a ${\beta}-amino$ acid, plays an important role as a neuromodulator and is necessary for the normal development of the brain. Since de novo synthesis of taurine in the brain is minimal and in vivo studies suggest that taurine dose not cross the blood-brain barrier, we examined whether the choroid plexus, the blood-cerebrospinal fluid (CSF) barrier, plays a role in taurine transport in the central nervous system. The uptake of $[^3H]-taurine$ into ATP depleted choroid plexus from rabbit was substantially greater in the presence of an inwardly directed $Na^+$ gradient taurine accumulation was negligible. A transient in side-negative potential gradient enhanced the $Na^+-driven$ uptake of taurine into the tissue slices, suggesting that the transport process is electrogenic, $Na^+-driven$ taurine uptake was saturable with an estimated $V_{max}$ of $111\;{\pm}\;20.2\;nmole/g/15\;min$ and a $K_M\;of\;99.8{\pm}29.9\;{\mu}M$. The estimated coupling ratio of $Na^+$ and taurine was $1.80\;{\pm}\;0.122.$ $Na^+-dependent$ taurine uptake was significantly inhibited by ${\beta}-amino$ acids, but not by ${\alpha}-amino$ acids, indicating that the transporter is selective for ${\beta}-amino$ acids. Since it is known that the physiological concentration of taurine in the CSF is lower than that in the plasma, the active transport system we characterized may face the brush border (i.e., CSF facing) side of the choroid plexus and actively transport taurine out of the CSF. Therefore, we examined in vivo elimination of taurine from the CSF in the rat to determine whether elimination kinetics of taurine from the CSF is consistent with the in vitro study. Using a stereotaxic device, cannulaes were placed into the lateral ventricle and the cisterna magna of the rat. Radio-labelled taurine and inulin (a marker of CSF flow) were injected into the lateral ventricle, and the concentrations of the labelled compounds in the CSF were monitored for upto 3 hrs in the cisterna magna. The apparent clearance of taurine from CSF was greater than the estimated CSF flow (p<0.005) indicating that there is a clearance process in addition to the CSF flow. Taurine distribution into the choroid plexus was at least 10 fold higher than that found in other brain areas (e. g., cerebellum, olfactory bulb and cortex). When unlabelled taurine was co-administered with radio-labelled taurine, the apparent clearance of taurine was reduced (p<0.0l), suggesting a saturable disposition of taurine from CSF. Distribution of taurine into the choroid plexus, cerebellum, olfactory bulb and cortex was similarly diminished, indicating that the saturable uptake of taurine into these tissues is responsible for the non-linear disposition. A pharmacokinetic model involving first order elimination and saturable distribution described these data adequately. The Michaelis-Menten rate constant estimated from in vivo elimination study is similar to that obtained in the in vitro uptake experiment. Collectively, our results demonstrate that taurine is transported in the choroid plexus via a $Na^+-dependent,saturable$ and apparently ${\beta}-amino$ acid selective mechanism. This process may be functionally relevant to taurine homeostasis in the brain.

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Draft Genome Sequence of the Yeast Strain Hormonema macrosporum POB-4, which Produces the Biosurfactant Glycocholic Acid

  • Parthiban Subramanian;Jeong-Seon Kim;Jun Heo;Yiseul Kim
    • Microbiology and Biotechnology Letters
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    • v.51 no.4
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    • pp.531-534
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    • 2023
  • We report the draft genome sequence of the yeast strain Hormonema macrosporum POB-4, capable of producing the biosurfactant glycocholic acid, one of the bile acids. A majority of genes with known function were associated with metabolism and transport of amino acid and carbohydrate as well as secondary metabolites biosynthesis, transport, and catabolism. We observed genes of eleven C-N hydrolases and two CoA transferases which have been reported to be involved in the biosynthesis of glycocholic acid. Further experimental studies can help to elucidate the specific genes responsible for biosurfactant production in strain POB-4.

Functional Characteristics of Neutral Amino Acid Transporter in Opossum Kidney (OK) Cells

  • Woo, Jae-Suk;Park, Moon-Hwan;Oh, Sae-Ok;Jung, Jin-Sup;Kim, Yong-Keun;Lee, Sang-Ho
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.2
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    • pp.185-193
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    • 1997
  • The characteristics of $Na^+$-dependent cycloleucine uptake was investigated in OK cells with regard to substrate specificity and regulation by protein kinase C (PKC). Inhibition studies with different synthetic and natural amino acids showed a broad spectrum affinity to neutral amino acids regardless of their different side chains including branched or aromatic, indicating that the $Na^+$-dependent cycloleucine uptake in OK cells is mediated by System $B^o$ or System $B^o$-like transporter rather than the classical System A or ASC. Phorbol 12-myristate 13-acetate (PMA) and phorbol 12,13-dibutyrate, but not $4{\alpha}-PMA$ elicited a time-dependent biphasic stimulation of $Na^+$-dependent cycloleucine uptake, which produced early transient peak at 30 min and late sustained peak at 180min. Both the early and late stimulations by PMA were due to an increase in Vmax and not due to a change in Km. PKC inhibitors blocked both the early and late stimulation by PMA, while protein synthesis inhibitors blocked the late stimulation only. These results suggest the existence and regulation by PKC of System $B^o$ or System $B^o$-like broad spectrum transport system for neutral amino acids in OK cells.

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Determination of Amino Acids on Wintertime PM2.5 using HPLC-FLD (HPLC-FLD를 이용한 겨울철 PM2.5 중 아미노산 성분 분석)

  • Park, Da-Jeong;Cho, In-Hwan;Bae, Min-Suk
    • Journal of Korean Society for Atmospheric Environment
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    • v.31 no.5
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    • pp.482-492
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    • 2015
  • Ground-based measurements were conducted from January 6 to 12 of 2015 for understanding characteristics of nitrogen containing carbonaceous aerosols as 16 amino acids at the Mokpo National University, Korea. The detailed amino acid components such as Cystine ($(SCH_2CH(NH_2)CO_2H)_2$) and Methionine ($C_5H_{11}NO_2S$) and their sources were analyzed by High-Performance Liquid Chromatography with Fluorescence Detection (HPLC-FLD) for behavior of secondary products in particulate matter. In addition, organic carbon (OC) and elemental carbon (EC) based on the carbonaceous thermal distribution (CTD), which provides detailed carbon signature characteristics relative to analytical temperature, and water soluble organic carbon (WSOC) by total organic carbon (TOC) analyzer were used to understand the carbon compound behaviors. The backward trajectories were discussed for originations of carbonaceous aerosols as well. Different airmasses were classified with the amino acids and OC thermal signatures. The results can provide to understand the aging process influenced by the long-range transport from East Sea area.

EFFECT OF INSULIN ADMINISTRATION ON THE COLLAGEN BIOSYNTHESIS IN INCISOR PULP AND GINGIVA OF RAT (Insulin 투여가 백서치수 및 치은의 Collagen 생합성에 미치는 영향)

  • Kim, Gwan-Shik
    • The Journal of the Korean dental association
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    • v.19 no.6 s.145
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    • pp.529-533
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    • 1981
  • The effect of insulin on the collagen metabolism in dental pulp and gingiva was studied using Sprague-Dawley rat and ³H-proline was injected intraperitoneally and then sacrificed according to predetermined schedule. Lower incisor pulp and gingiva were removed immediatedly and hydrolysed with acid. After digestion, proline and hydroxyproline were separated and analysed quantitatively employing thin-layer chromatography and radioactivity of each amino acid was counted by liquid scintillation counter. The label incorporated into lower incisor pulp and gingiva and radioactivity of ³H-hydroxyproline were increased in insulin administered group. And disappearance rate of ³H-hydroxyproline was decreased. Those results indicated that amino acid, I.e. proline transport and collagen biosynthesis were increased in dental pulp and gingiva by insulin administration.

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Gene Expression Profiling of Liver and Mammary Tissues of Lactating Dairy Cows

  • Baik, M.;Etchebarne, B.E.;Bong, J.;VandeHaar, M.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.6
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    • pp.871-884
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    • 2009
  • Gene expression profiling is a useful tool for identifying critical genes and pathways in metabolism. The objective of this study was to determine the major differences in the expression of genes associated with metabolism and metabolic regulation in liver and mammary tissues of lactating cows. We used the Michigan State University bovine metabolism (BMET) microarray; previously, we have designed a bovine metabolism-focused microarray containing known genes of metabolic interest using publicly available genomic internet database resources. This is a high-density array of 70mer oligonucleotides representing 2,349 bovine genes. The expression of 922 genes was different at p<0.05, and 398 genes (17%) were differentially expressed by two-fold or more with 222 higher in liver and 176 higher in mammary tissue. Gene ontology categories with a high percentage of genes more highly expressed in liver than mammary tissues included carbohydrate metabolism (glycolysis, glucoenogenesis, propanoate metabolism, butanoate metabolism, electron carrier and donor activity), lipid metabolism (fatty acid oxidation, chylomicron/lipid transport, bile acid metabolism, cholesterol metabolism, steroid metabolism, ketone body formation), and amino acid/nitrogen metabolism (amino acid biosynthetic process, amino acid catabolic process, urea cycle, and glutathione metabolic process). Categories with more genes highly expressed in mammary than liver tissue included amino acid and sugar transporters and MAPK, Wnt, and JAK-STAT signaling pathways. Real-time PCR analysis showed consistent results with those of microarray analysis for all 12 genes tested. In conclusion, microarray analyses clearly identified differential gene expression profiles between hepatic and mammary tissues that are consistent with the differences in metabolism of these two tissues. This study enables understanding of the molecular basis of metabolic adaptation of the liver and mammary gland during lactation in bovine species.

Effect on the Arginine Transport of Mutant MCAT1, Mouse Cationic Aminoacid Transporter (MCAT1의 돌연변이체가 Arginine 통과 능력에 미치는 영향)

  • Kim, Jung-Woo
    • The Journal of Natural Sciences
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    • v.8 no.2
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    • pp.35-41
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    • 1996
  • To find the substrate interacting site of the MCAT1, charged amino acid residues in the transmembrane domain were changed to opposite charged amino acids and studied the arginine uptake, gp70 binding, efflux and protein expression using the Xenopus oocyte expression method. Among the five mutants of MCAT1, the D403K showed the most interesting characteristics, which had normal gp70 binding but low arginine uptake function, that means the normal expression on the membrane but decreased transport function. All mutants except K211E showed decreased arginine efflux, and kinetic study showed decreased Vmax. Together, Clu(403) residue of MCAT1 may show the possible substrate interacting site in the transmembrane domain of MCAT1.

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Synthesis and In-vitro Evaluation of N4-Amino Acid Derivatives of Cytarabine for Improving the Oral Delivery of Cytarabine

  • Jin, Ming-Ji;Hong, Joon-Hee;Han, Hyo-Kyung
    • Journal of Pharmaceutical Investigation
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    • v.38 no.4
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    • pp.255-259
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    • 2008
  • The present study aimed to investigate the in-vitro characteristics of N4-amino acid derivatives of cytarabine for the oral delivery of cytarabine. After the synthesis of L-Ile-cytarabine, L-Leu-cytarabine and L-Arg-cytarabine, the gastrointestinal stability of each prodrug was examined using artificial gastric juice and intestinal fluids. The cellular uptake characteristics of prodrugs were also examined in Caco-2 cells. While L-Ile-cytarabine and L-Leu-cytarabine appeared to be stable in all the tested biological media during 4-hr incubation, L-Arg-cytarabine was rapidly disappeared within 5 min. Accordingly, the cellular uptake of L-Ile-cytarabine and L-Leu-cytarabine was significantly higher than that of its parent drug, cytarabine in Caco-2 cells but the cellular uptake of L-Arg-cytarabine was similar to that from its parent drug. The cellular uptake of L-Ile-cytarabine and L-Leu-cytarabine appeared to be saturable as drug concentration increased from 0.4 to 4 mM. Collectively, L-Ile-cytarabine and L-Leu-cytarabine could be promising candidates to improve the oral absorption of cytarabine via a saturable transport pathway.

Alterations in Membrane Transport Function and Cell Viability Induced by ATP Depletion in Primary Cultured Rabbit Renal Proximal Tubular Cells

  • Lee, Sung-Ju;Kwon, Chae-Hwa;Kim, Yong-Keun
    • The Korean Journal of Physiology and Pharmacology
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    • v.13 no.1
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    • pp.15-22
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    • 2009
  • This study was undertaken to elucidate the underlying mechanisms of ATP depletion-induced membrane transport dysfunction and cell death in renal proximal tubular cells. ATP depletion was induced by incubating cells with 2.5 mM potassium cyanide(KCN)/0.1 mM iodoacetic acid(IAA), and membrane transport function and cell viability were evaluated by measuring $Na^+$-dependent phosphate uptake and trypan blue exclusion, respectively. ATP depletion resulted in a decrease in $Na^+$-dependent phosphate uptake and cell viability in a time-dependent manner. ATP depletion inhibited $Na^+$-dependent phosphate uptake in cells, when treated with 2 mM ouabain, a $Na^+$ pump-specific inhibitor, suggesting that ATP depletion impairs membrane transport functional integrity. Alterations in $Na^+$-dependent phosphate uptake and cell viability induced by ATP depletion were prevented by the hydrogen peroxide scavenger such as catalase and the hydroxyl radical scavengers(dimethylthiourea and thiourea), and amino acids(glycine and alanine). ATP depletion caused arachidonic acid release and increased mRNA levels of cytosolic phospholipase $A_2(cPLA_2)$. The ATP depletion-dependent arachidonic acid release was inhibited by $cPLA_2$ specific inhibitor $AACOCF_3$. ATP depletion-induced alterations in $Na^+$-dependent phosphate uptake and cell viability were prevented by $AACOCF_3$. Inhibition of $Na^+$-dependent phosphate uptake by ATP depletion was prevented by antipain and leupetin, serine/cysteine protease inhibitors, whereas ATP depletion-induced cell death was not altered by these agents. These results indicate that ATP depletion-induced alterations in membrane transport function and cell viability are due to reactive oxygen species generation and $cPLA_2$ activation in renal proximal tubular cells. In addition, the present data suggest that serine/cysteine proteases play an important role in membrane transport dysfunction, but not cell death, induced by ATP depletion.