• Title/Summary/Keyword: Palmitoyl-CoA

Search Result 22, Processing Time 0.025 seconds

Roles of cysteine residues in the inhibition of human glutamate dehydrogenase by palmitoyl-CoA

  • Son, Hyo Jeong;Ha, Seung Cheol;Hwang, Eun Young;Kim, Eun-A;Ahn, Jee-Yin;Choi, Soo Young;Cho, Sung-Woo
    • BMB Reports
    • /
    • v.45 no.12
    • /
    • pp.707-712
    • /
    • 2012
  • Human glutamate dehydrogenase isozymes (hGDH1 and hGDH2) have been known to be inhibited by palmitoyl-CoA with a high affinity. In this study, we have performed the cassette mutagenesis at six different Cys residues (Cys59, Cys93, Cys119, Cys201, Cys274, and Cys323) to identify palmitoyl-CoA binding sites within hGDH2. Four cysteine residues at positions of C59, C93, C201, or C274 may be involved, at least in part, in the inhibition of hGDH2 by palmitoyl-CoA. There was a biphasic relationship, depending on the levels of palmitoyl-CoA, between the binding of palmitoyl-CoA and the loss of enzyme activity during the inactivation process. The inhibition of hGDH2 by palmitoyl-CoA was not affected by the allosteric inhibitor GTP. Multiple mutagenesis studies on the hGDH2 are in progress to identify the amino acid residues fully responsible for the inhibition by palmitoyl-CoA.

MEASUREMENT OF SYNTHESIS RATE OF LONG-CHAIN ACYL-COENZYME A ESTER IN BOVINE LIVER BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY

  • Mitsuhashi, T.;Mitsumoto, M.;Yamashita, Y.;Ozawa, S.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.1 no.2
    • /
    • pp.99-106
    • /
    • 1988
  • A high performance liquid chromatographic procedure is described for the direct determination of the picomole amount of palmitoyl-Coenzyme A and stearoyl-Coenzyme A, using a stainless steel column packed with C-18 derivatized porous silica ($5{\mu}m$), an isocratic elution with a mixture of 33 mM $KH_2PO_4$/acetonitrile as a mobile phase and a UV detector. The long-chain acyl-Coenzyme A esters were determined in incubated microsomal fractions of a bovine liver to demonstrate the utility of this method for monitoring acyl-CoA synthesis in biological samples. The reaction rate of palmitate was higher than that of stearate. After a 60 minute incubation period, the generated amount of palmitoyl-Coenzyme A and stearoyl-Coenzyme A were approximately 70 and 20 n mol/mg micresomal protein, respectively. The advantage of this method are in that no decomposition of the CoA esters is involved, while the constituent molecular species is detected.

Biochemical Characteristics of a Palmitoyl Acyl Carrier Protein Thioesterase Purified from Iris pseudoacorus

  • Kang, Han-Chul;Hwang, Young-Soo
    • BMB Reports
    • /
    • v.29 no.5
    • /
    • pp.436-441
    • /
    • 1996
  • The palmitoyl acyl carrier protein (ACP) specific thioesterase (EC 3.1.2.14) from Iris pseudoacorus was purified and characterized. The thioesterase which was very unstable in relatively high salt concentrations was eluted using a co-gradient of Triton X-100 and low concentration of KCl or Na-phosphate from Q-Sepharose, DEAE-Sepharose, and hydroxyapatite chromatography. SDS-PAGE analysis showed a single band with a molecular weight of 35,000. The native molecular weight of approximately 37,000 was estimated by Sephacryl S-200 chromatography, indicating that the enzyme is a monomer. The thioesterase activity was inhibited about 75% and 50% by N-ethylmaleimide (2 mM) and phenylmethylsulfonyl fluoride (2 mM). respectively. The N-ethylmaleimide-inactivation was protected by sodium palmitate but the inactivation with phenylmethylsulfonyl fluoride was not protected. Oxidation of thiols by 2 mM 5.5'-dithio-bis-(2-nitrobenzoic acid) resulted in 65% inactivation of the enzyme. These results suggest that a cysteinyl residue is essential to the catalytic reaction of the enzyme. The enzyme activity was increased by sodium citrate and also by $Cu^{2+}$

  • PDF

L-Carnitine Reduces Obesity Caused by High-Fat Diet in C57BL/6J Mice

  • Mun, Eun-Gyeng;Soh, Ju-Ryoun;Cha, Youn-Soo
    • Food Science and Biotechnology
    • /
    • v.16 no.2
    • /
    • pp.228-233
    • /
    • 2007
  • This study evaluated the effects of carnitine supplementation on obesity caused by a high-fat diet in C57BL/6J mice. The mice were fed a normal diet (ND), high-fat diet (HD), or carnitine-supplemented (0.5% of diet) high-fat diet (HDC) for 12 weeks. The results showed that body weight, energy intake, and feed intake were lower in the HDC group than the control groups. Acid-soluble acylcarnitine (A SAC), acid-insoluble acylcarnitine (AIAC), and total carnitine (TCNE) in the serum and liver were significantly higher in the HDC group. Hepatic carnitine palmitoyl transferase-I activity was significantly higher in the HDC group than the control groups. Acyl-coA synthetase (ACS) and carnitine palmitoyl transferase-I (CPT-I) mRNA expression in the liver was highest in the HDC group, however hepatic acetyl-coA carboxylase (ACC) mRNA expression in this group was lowest. Serum leptin levels and abdominal fat weight were lowest in the HDC group. We concluded that L-carnitine supplementation diminished the risk of obesity caused by a high-fat diet.

Cell Biological Studies on the Mechanism of Development and Differentiation Ⅷ 3. Effects of Polyamines on the Activities of Corn Glucose-6-Phosphate Dehydrogenase, 6-Phosphogluconate Dehydrogenase, and Protein Kinase (생체발생 및 분화기구의 세포생물학적 연구 Ⅷ 3. Polyamine이 옥수수 배의 Glucose-6-Phosphate Dehydrogenase, 6-Phosphogluconate Dehydrogenase 및 Protein Kinase 활성에 미치는 영향)

  • Cho, Young Dong
    • Journal of Plant Biology
    • /
    • v.27 no.2
    • /
    • pp.73-80
    • /
    • 1984
  • Palmitoyl CoA was found to inhibit corn embryo axis glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase, which were also inhibites by polyamines. However, reversal of inhibition of both enzymes by palmitoyl CoA was made by spermine. Activity of corn embryo axis protein kinase was found to increase steadily after germination. Activation and inhibition of protein kinase were made by MgCl$_2$and all polymines, respectively. Suc results suggest that fatty acid biosynthesis and lypolysis could be regulated to some extent by polyamines in corn embryo axis.

  • PDF

Antimicrobial Activities of Ethosome-Encapsulated Palmitoyl Tripeptide (Ethosome에 캡슐화된 Palmitoyl Tripeptide의 항균효과)

  • Lee, Yeon-Jung;Lee, Yun Sub;Jin, Byung Suk
    • Applied Chemistry for Engineering
    • /
    • v.25 no.6
    • /
    • pp.570-576
    • /
    • 2014
  • Palmitoyl tripeptide (M330) showed higher antimicrobial activities than methyl paraben or phenoxy ethanol through minimum inhibitory concentration (MIC) test. However, when the M330 was added into cosmetic formulation, white precipitates formed due to the electrostatic interaction between M330 and carbopol (carboxy vinyl polymer) as a thickener in cosmetics, and the viscosity of cosmetics decreased sharply. Also, the antimicrobial activities of M330 in cosmetics became lower than those of methyl paraben or phenoxy ethanol. Thus, the encapsulation of M330 in ethosome vesicle was attempted in order to recover the declined antimicrobial activities of M330 in cosmetics and prevent the precipitates from forming. When ethosome-encapsulated M330 was added into cosmetics, the precipitates did not form, and the decrease in the viscosity of cosmetics was not large compared to the addition of unencapsulated M330. Challenge tests showed that antimicrobial activities against gram negative bacteria were improved by the encapsulation of M330, but the encapsulation was not effective against gram positive bacteria and fungus. A combination of M330 with EDTA showed synergistic inhibitory potential against C. albicans. After coencapsulation of M330 and EDTA in ethosome, antimicrobial activities proved to be higher than those of unencapsulated M330 and EDTA.

Studies on the Development of Cocoa Butter Equivalent Fat by Reverse - Micelle Enzyme Reaction System (역마이셀-효소반응계에 의한 코코아 버터 대용지 개발에 관한 연구)

  • Yoon, Seung-Heon;Shin, Woong-Kyu;Lee, Yoon-Hyung;Rhee, Kyu-Soon
    • Korean Journal of Food Science and Technology
    • /
    • v.24 no.2
    • /
    • pp.111-116
    • /
    • 1992
  • Production of cocoa butter equivalent fat (CBE) from palm oil and stearic acid by reverse micelle lipase reaction system was studied. Qualitative and quantitative analyses of triglycerides were performed by HPLC. The reaction conditions for maximum conversion from triolein and stearic acid to 1-stearoyl-2,3-dioleoyl glycerol(SOO) and 1,3-distearoyl-2-oleoyl glycerol(SOS) were as follows: a molar ratio of water/Aerosol OT, 10; triolein, 30 mM; stearic acid, 90 mM; pH, 7.5; and temperature, $50^{\circ}C$. By lipase in reverse micellar system containing palm oil and stearic acid, 1,3-dipalmitoyl-2-oleoyl glycerol(POP), 1-palmitoyl-2,3-dioleoyl glycerol(POO) and SOO decreased, but large amounts of 1-palmitoyl-2-oleoyl-3-stearoyl glycerol(POS) and SOS was formed.

  • PDF

A Monoacyldigalactosyl Glycerol from the Green Alga Enteromorpha prolifera

  • Kim, Ju-Sun;Shim, Sang-Hee;Lee, Sang-Hyun;Chae, Sung-Wook;Han, Sang-Jun;Kang, Sam-Sik;Lee, Yeon-Sil;Jung, Sang-Hoon;Shin, Kuk-Hyun
    • Natural Product Sciences
    • /
    • v.10 no.6
    • /
    • pp.341-343
    • /
    • 2004
  • A monoacyldigalactosyl glycerol was isolated from the $CH_2Cl_2$ soluble fraction of the MeOH extract from the green alga Enteromorpha prolifera. The structure was established as $1-O-palmitoyl-3-O-[{\alpha}-D-galactopyranosyl(1{\rightarrow}6)-{\beta}-D-galactopyranosyl]-sn-glycerol$ (1) by chemical and spectroscopic methods.

1-palmitoyl-2-linoleoyl-3-acetyl-rac-glycerol (EC-18) Modulates Th2 Immunity through Attenuation of IL-4 Expression

  • Yoon, Sun Young;Kang, Ho Bum;Ko, Young-Eun;Shin, Su-Hyun;Kim, Young-Jun;Sohn, Ki-Young;Han, Yong-Hae;Chong, Saeho;Kim, Jae Wha
    • IMMUNE NETWORK
    • /
    • v.15 no.2
    • /
    • pp.100-109
    • /
    • 2015
  • Controlling balance between T-helper type 1 (Th1) and T-helper type 2 (Th2) plays a pivotal role in maintaining the biological rhythm of Th1/Th2 and circumventing diseases caused by Th1/Th2 imbalance. Interleukin 4 (IL-4) is a Th2-type cytokine and often associated with hypersensitivity-related diseases such as atopic dermatitis and allergies when overexpressed. In this study, we have tried to elucidate the function of 1-palmitoyl-2-linoleoyl-3-acetyl-rac-glycerol (EC-18) as an essential modulator of Th1/Th2 balance. EC-18 has showed an inhibitory effect on the production of IL-4 in a dose-dependent manner. RT-PCR analysis has proved EC-18 affect the transcription of IL-4. By analyzing the phosphorylation status of Signal transducer and activator of transcription 6 (STAT6), which is a transcriptional activator of IL-4 expression, we discovered that EC-18 induced the decrease of STAT6 activity in several stimulated cell lines, which was also showed in STAT6 reporter analysis. Co-treatment of EC-18 significantly weakened atopy-like phenotypes in mice treated with an allergen. Collectively, our results suggest that EC-18 is a potent Th2 modulating factor by regulating the transcription of IL-4 via STAT6 modulation, and could be developed for immune-modulatory therapeutics.

Glycerides from the Aerial Parts of Garland (Chrysanthemum coronarium L.) and Their Inhibitory Effects on ACAT, DGAT, FPTase, and $\beta$-Secretase

  • Song, Myoung-Chong;Yang, Hye-Joung;Cho, Jin-Gyeong;Chung, In-Sik;Kwon, Byoung-Mog;Kim, Dae-Keun;Baek, Nam-In
    • Food Science and Biotechnology
    • /
    • v.18 no.1
    • /
    • pp.95-102
    • /
    • 2009
  • The aerial parts of garland (Chrysanthemum coronarium L.) were extracted in 80% aqueous methanol (MeOH) and the concentrated extract was then partitioned using ethyl acetate (EtOAc), n-butanol (n-BuOH), and $H_2O$, successively. EtOAc and n-BuOH fractions resulted in 4 glycerides with the application of octadecyl silica gel and silica gel column chromatography. The chemical structures of the glycerides were determined using several spectroscopic methods, including nuclear magnetic resonance (NMR) and mass spectrometry (MS) as (2S)-1-O-palmitoyl-sn-glycerol (1), (2S)-1-O-oleoyl-2-O-oleoyl- 3-O-$\beta$-D-galactopyranosyl-sn-glycerol (2), (2S)-1-O-palmitoyl-2-O-linoleoyl-3-O-phosphorouscholine-sn-glycerol (3), and (2S)-1-O-linolenoyl-2-O-palmitoyl-3-O-[$\alpha$-D-galactopyrasyl-($1{\rightarrow}6$)-$\beta$-D-galactopyranosyl]-sn-glycerol (4). The free fatty acids of these glycerides were determined with gas chromatography (GC)-MS analysis following alkaline hydrolysis and methylation. These glycerides demonstrated an inhibitory effect on acyl-CoA: cholesterol acyltransferase (ACAT, compound 1: $45.6{\pm}0.2%$ at $100{\mu}g/mL$), diacylglycerol acyltransferase (DGAT, compound 1: $59.1{\pm}0.1%$ at $25{\mu}g/mL$), farnesyl protein transferase (FPTase, compound 2: $98.0{\pm}0.1%$; compound 3: $55.2{\pm}0.1%$ at $100{\mu}g/mL$), and $\beta$-secretase ($IC_{50}$, compound 4: $2.6{\mu}g/mL$) activity. This paper is the first report on the isolation of these glycerides from garland and their inhibitory activity on ACAT, DGAT, FPTase, and $\beta$-secretase.