• Title/Summary/Keyword: protein binding

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Characterization of an Ion Channel Prepared from Tomato Roots and Inhibitory Effects by Heavy Metal Ions (토마토 뿌리조직에서 분리한 이온채널의 중금속에 의한 저해)

  • Shin, Dae-Seop;Han, Min-Woo;Kim, Young-Kee
    • Applied Biological Chemistry
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    • v.47 no.4
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    • pp.390-395
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    • 2004
  • In order to characterize ion channels present in tomato roots, microsomes were incorporated into an artificial lipid bilayer arranged for electrophysiological analysis. Of the five different ion channels that could be found, a channel of 450 pS conductance was found most frequently. This channel displayed subconductance states of 450, 257 and 105 pS. All subconductance states showed linear current-voltage relationships. At positive holding potentials, high frequency of transient channel openings was observed; however, at negative potentials, the open times were long and open probability high. Po was 0.83 at -40 mV. When an additional 50 mM $K^+\;or\;Na^+$ was added to the cis side of bilayer, the reversal potentials shifted in the negative direction to near -10 mV. Thus, the 450 pS cation channel selects poorly between $K^+\;and\;Na^+$. In the presence of $100\;{\mu}M$ metal ions, the channel activity was severely inhibited by $La^{3+},\;Ba^{2+},\;and\;Zn^{2+}$, and Po was decreased to 0.2 or even less. However, $Al^{3+}\;and\;Cd^{2+}$ decreased the activity by only 20%. Interestingly, each metal ion showed different kinetics of channel inhibition. While $500\;{\mu}M\;La^{3+}$ inhibited the activities of all subconductance state, 1 mM $Zn^{2+}$ inhibited all except the 105 pS state. $Cd^{2+}$ changed the gating of the channel from a long-opening state to brief transient openings even at negative holding potentials. These data represent that the metal ions may have different binding sites on the channel protein and could be useful modulators and probes to investigate structural characteristics as well as the functional roles of the 450 pS channel on the root physiology.

Polymorphisms of TAS1R3 and GNAT3 Genes Are Associated with Patients with Taste Disorder (미각장애와 TAS1R3 및 GNAT3 유전자의 다형성과의 연관성)

  • Bae, Jae-Woong;Kim, Un-Kyung;Kwon, Tae-Jun;Choi, Su-Jin;Ye, Mi-Kyung
    • Journal of Life Science
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    • v.21 no.3
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    • pp.412-416
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    • 2011
  • Taste sensation plays a crucial role in selecting and ingesting foods with different qualities which convey information about their nutrient content and/or safety. Sweetness is one of the five modalities in humans and serves as an energy resource for metabolism. There are reports on allelic polymorphisms which influence perception of sweetness in mice and humans. Since the influence of genetic factors on taste disorder has not been studied, we investigated the association of genetic polymorphisms in TAS1R3 and guanine nucleotide binding protein, alpha transducing 3 (GNAT3) genes and taste disorder. A total of 150 individuals composed of 50 patients with taste disorder and 100 healthy controls were recruited for the study and PCR-mediated directing sequencing method was used to genotype for two different single nucleotide polymorphisms (SNPs) - rs307355 (T>C) and rs35744813 (T>C) in the TAS1R3 gene, and rs7792845 (T>C) and rs1524600 (C>T) in the the GNAT3 gene. The allele and genotype frequencies of rs307355 and rs35744813 in the TAS1R3 gene showed a significant association between patients with taste disorder (p=0.022 and p=0.013 in both of SNPs, respectively). In addition, the frequency of T-T haplotype in the TAS1R3 gene was higher in taste disorder cases than in the controls (OR, 1.93: 95%. CI, 1.09-3.39, p=0.022). In the GNAT3, the genotype frequency of rs7792845 in the patients was also different from the controls (p=0.048), but allele frequency was not significantly associated in either group. Our result provides the frequencies of SNPs and haplotypes of the TAS1R3 and GNAT3 genes for the fundamental information of nutrigenetics in perception of the taste of sweetness in the Korean population. Also, the study suggests that the allelic polymorphisms of TAS1R3 and GNAT3 genes may be useful as a molecular marker for evaluating patients with taste disorder. Further studies with large samples are required to clarify our observation.

Anti-obesity Effects of Peucedanum japonicum Thunberg L. on 3T3-L1 Cells and High-fat Diet-induced Obese Mice (식방풍잎(Peucedanum japonicum Thunberg L.)의 물추출물이 3T3-L1 세포와 고지방식이로 유도된 마우스에서 항비만 효과)

  • Jung, Ho-Kyung;Sim, Mi-Ok;Jang, Ji-Hun;Kim, Tae-Muk;An, Byeong-Kwan;Kim, Min-Suk;Jung, Won Seok
    • Korean Journal of Plant Resources
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    • v.29 no.1
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    • pp.1-10
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    • 2016
  • Obesity is a pro-inflammatory state that contributes to the development of metabolic disorders such as hyperlipidemia, insulin resistance, type 2 diabetes, non-alcoholic fatty liver, and cardiovascular disease. In this study, we evaluated the inhibition of adipogenesis in 3T3-L1 cells and in high-fat diet (HFD)-induced obese mice by Peucedanum japonicum Thunberg L. water extract (PJT). Lipid accumulation measurement indicates that PJT markedly inhibited adipogenesis in a dose-dependent manner. RT-PCR results demonstrated that the mRNA expression of adipogenic transcription factors such as peroxisome proliferator-activated receptor-γ (PPARγ) and CCAAT/enhancer binding protein-α (C/EBPα) in 3T3-L1 cells were significantly down-regulated by PJT treatment. Oral administration of PJT (100, 300, and 500 ㎎/㎏, b.w/daily for 4 weeks) was conducted in high-fat diet induced obese mice and C57BL/6 mice. The PJT-administered group of HFD-induced mice had a lower body weight gain, along with decreased serum levels of glucose, triglycerides, and total cholesterol compared with the control mice, however, the HDL-cholesterol/total cholesterol ratio was increased. Furthermore, the elevated mRNA expression levels of adipogenesis related genes in the white adipose tissue of obese mice were significantly suppressed by PJT. These results indicate that PJT exhibits anti-obesity effects in obese mice by decreasing in serum lipid levels and lipogenesis related gene.

Enzymatic Characteristics of Water-Insoluble ${\alpha}-Amylase$ Immobilized on Dithiocarbamate Wool (Dithiocarbamate Wool에 고정(固定)된 불용성(不溶性) ${\alpha}-Amylase$의 특성(特性)에 대하여)

  • Lee, Kyung-Hee;Kim, Jong-Deog;Lee, Kang-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.14 no.2
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    • pp.164-170
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    • 1985
  • Yellowish modified wool, dithiocarbamate(DTC) wool, was synthesized by partial hydrolysis in 0.2 N-NaOH reacting with carbon disulfide to use as ${\alpha}-amylase$ immobilization matrix. ${\alpha}-amylase$ was immobilized reacting with sulfide group of DTC-wool by covalent binding within 1 hour. 0.5 gram of this preparation, $DTC-wool-{\alpha}-amylase$, contained 150 ug of enzyme protein and its specific activity was about 90% of the native one. General properties of $DTC-wool-{\alpha}-amylase$ were a little different from optimum temperature, optimum pH, heat stability, kinetic constants and activation energy. An apparent Michaelis constant and maximum velocity of $DTC-wool-{\alpha}-amylase$ were 5.56 mg/ml and 0.37 mg/ml. $min^{-1}$ respectively, while activation energy was 16.6 kcal/mole.

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Quality Characteristics of Adzuki Beans Sediment According to Variety (품종에 따른 팥 앙금의 품질 특성)

  • Song, Seuk-Bo;Seo, Hye-In;Ko, Jee-Yeon;Lee, Jae-Saeng;Kang, Jong-Rae;Oh, Byeong-Geun;Seo, Myung-Chul;Yoon, Young-Nam;Kwak, Do-Yeon;Nam, Min-Hee;Woo, Koan-Sik
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.8
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    • pp.1121-1127
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    • 2011
  • We evaluated the quality characteristics of adzuki bean sediment according to variety. The moisture, crude protein, and crude ash contents of the various adzuki bean varieties were 8.2~11.1, 15.4~20.6 and 3.3~3.6 g/100 g, respectively. The potassium contents of Chilbo-pat (CB) and Hongeon-pat (HE) were 875.1 and 873.1 mg/100 g, respectively. The calcium contents of Jungbu-pat (JB) and Kumsil-pat (KS)were 73.6 and 73.2 mg/100 g, respectively. A high level of magnesium (131.4 mg/100 g) was found in Yeonkeum-pat (YK). The yields of adzuki bean sediment according to variety were no different either wet (188.3~204.7%) or dry (62.1~66.0%). The L-values on sediment of YK and KS were 67.0 and 68.0, respectively; however, the CB L-value was low at 54.0. A high level of a- (6.6) and b-value (12.8) was found in YK; however, the values for CB were much lower at 3.8 and 5.9, respectively. There was no difference in particle-size distribution, water binding capacity, and solubility of adzuki bean sediment according to variety. High levels of peak (3.79 RVU), trough (3.75 RVU), final (7.33 RVU), and setback viscosity (3.54 RVU) were found in JB. The sensory properties of products in food processing are important, and the variety of adzuki bean sediment should be chosen depending on desired product characteristics.

Molecular Phylogenetic Study of the Endangered Land Snail Satsuma myomphala Based on Metallothionein Gene. (Metallothionein 유전자를 기초로 한 멸종위기 육상 달팽이 Satsuma myomphala (거제외줄달팽이) 의 분자계통학적 연구)

  • Sang, Min Kyu;Kang, Se Won;Hwang, Hee-Ju;Chung, Jong Min;Song, Dae Kwon;Min, Hye Rin;Park, Jie Eun;Ha, Hee Cheol;Lee, Hyun Jun;Hong, Chan Eui;Ahn, Young Mo;Park, So Young;Park, Young-Su;Park, Hong Seog;Han, Yeon Soo;Lee, Jun Sang;Lee, Yong Seok
    • The Korean Journal of Malacology
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    • v.32 no.4
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    • pp.263-268
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    • 2016
  • Metallothionein (MT) family of metal-binding proteins are involved in maintaining homeostasis and heavy metal poisoning. Recently, MT has been considered as a biomarker that can identify a particular species, very similar to the use of cytochrome oxidase I (COI) gene. Satsuma myomphala species of land snails have been reported from North-East Asia, including South Korea and Japan. In particular, the land snail species have been known from only a limited area of Geoje Island, Gyeongsangnam-do province of South Korea. Genetic studies of S. myomphala has been limited with only 6 nucleotide, 2 protein registered on the NCBI server. For elucidating the genetic information of S. myomphala, we conducted RNA sequencing analysis using Illumina HiSeq 2500 next-generation platform. We screened the MT gene from the RNA-Seq database to confirm the molecular phylogenetic relationship. After sequencing, the de novo analysis and clustering generated 103,774 unigenes. After annotation against PANM database using BLAST program, we obtained MT sequence of 74 amino acid residues containing the coding region of 222 bp. Based on this sequence, we found about 53 sequences using the BLAST program in NCBI nr database. Using ClustalX alignment, Maximum-Likehood Tree of MEGA program, we confirmed the molecular phylogenetic relationships that showed similarity with mollusks such as Helix pomatia and H. aspersa, Megathura crenulata.

Bioaccumulation of Heavy Metals in Ruditapes philippinarum (바지락 (Ruditapes philippinarum) 의 중금속 축적에 관한 연구)

  • Lee, Yong-Seok;Jo, Yong-Hun;Byun, In-Sun;Kang, Se-Won;Cho, Eun-Mi;Han, Yeon-Soo;Choi, Sang-Haeng;Park, Hong-Seog;Kho, Weon-Gyu;Ahn, In-Young;Jeong, Kye-Heon
    • The Korean Journal of Malacology
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    • v.22 no.2
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    • pp.157-165
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    • 2006
  • The present study was conducted to confirm that a bivalve Ruditapes philippinarum can be used as a biomarker for the monitoring of the heavy metal pollution in the silt of the marine environment. The clams were collected from the silt of Cheonsu-bay, Buheung-ri, and Tan-island of the West Sea, Korea. To observe the normal structures of the target organs (hepatopancreas and gill), they were dissected out for the immunohistochemical study and the electron microscopy with TEM, SEM, and SEM-EDS. The immunohistochemical study showed that the interdiverticular connective tissues of the hepatopancreas, and the outer epithelium of the gill lamellae was strongly reacted to anti-metallothionein (MT), indicating the presence of MT, a metal-binding protein, involved in metal detoxifying process. According to the examinations under the TEM, the epithelial cells of the hepatopancreas of the clams collected from polluted area (Tan-island) showed certain changes such as swollen rER, swollen nuclear envelope and inclusion bodies in the nulcei. In the SEM-EDS analysis, tissue of the hepatopancreas showed relatively higher concentration of S, Zn, and Cd. These elements are supposed to be concerning with the MT-reaction in the hepatopancreas. Considering that the coastal bivalve R. philippinarum showed immediate subcellular responses to heavy metal pollution in the overall experiments conducted, this species might act as one of efficient biomarkers for the heavy metal contamination in the marine environment.

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A Novel Glycine-Rich Region in Sox4 is a Target for the Proteolytic Cleavage in E. coli (전사활성 인자인 Sox4의 단백질 분해효소에 의한 표적 부위에 관한 연구)

  • 허은혜;최주연;장경희;김인경;임향숙
    • Korean Journal of Microbiology
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    • v.38 no.3
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    • pp.153-161
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    • 2002
  • Sox4, a transcription factor, consists of three functional domains: an HMG-box domain as a DNA binding domain, serine rich region as a transactivation domain and glycine rich region (GRR), an unknown functional domain. Although Sox4 is known to be functionally involved in heart, B-cell and reproductive system development, its physiological function remains to be elucidated. We used pGEX expression system to develop a simple and rapid method for purifying Sox4 protein in suitable forms for biochemical studies of their functions. Unexpectedly, we observed that full-length Sox4 appears to be protease-sensitive during expression and purification in E. coli. To map the protease-sensitive site in Sox4, we generated various constructs with each of functional domains of Sox4 and purified as the GST-Sox4 fusion proteins using glutathione beads. We found that the specific cleavage site for the proteolytic enzyme, which exists in E. coli, is localized within the novel GRR of Sox4. Our study suggest that the GRR of Sox4 may a target for the cellular protease action and this cleavage in the GRR may be involved in regulating physiological function of Sox4. Additionally, our study may provide a useful method for investigating the proteolytic cleavage of the target molecule in E. coli.

Inhibition of Pancreatic Lipase Activity and Adipocyte Differentiation in 3T3-L1 Cells Treated with Purple Corn Husk and Cob Extracts (자색옥수수 포엽과 속대 추출물의 리파아제 저해활성 및 3T3-L1 지방전구세포에서의 지방분화 억제효과)

  • Lee, Ki Yeon;Hong, Soo Young;Kim, Tae Hee;Kim, Jai Eun;Park, A-Reum;Noh, Hee Sun;Kim, Si Chang;Park, Jong Yeol;Ahn, Mun Seob;Jeong, Won Jin;Kim, Hee Yeon
    • Journal of Food Hygiene and Safety
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    • v.33 no.2
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    • pp.131-139
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    • 2018
  • Our review begins with the maize hybrid for grain, called 'Seakso 1,' which was developed in 2008 by the Gangwon Agricultural Research and Extension Services in Korea, and subsequently registered in 2011. In this study, we aimed to investigate the lipid metabolic enzyme activity and inhibitory effect on the adipocyte differentiation, in 3T3-L1 cells of the identified Seakso 1 corn husk and cob extracts (EHCS). We investigated the pancreatic lipase inhibitory effect and anti-adipogenic effect of EHCS.The lipid accumulation and adipocyte differentiation were measured by the procedure of Oil Red O staining, Real-time PCR and the Western blot analysis. The pancreatic lipase inhibitory activity of EHCS was measured at higher levels than those of the positive control (orlistat) at 100, 500, and $1,000{\mu}g/mL$. In particular, EHCS was noted as being significantly inhibited and including a measured adipocyte differentiation and lipid accumulation, when treated during the adipocyte differentiation process in 3T3-L1 cells. Based on the Oil Red O staining, EHCS inhibited lipid accumulation at 19.19%, 33.30% at $1000{\mu}g/mL$, $2000{\mu}g/mL$, respectively. The real-time PCR and Western blot analysis showed that EHCS significantly decreased in the mRNA expression and protein level of obesity-related factors, such as peroxisome-proliferatorsactivated-receptor-${\gamma}$ ($PPAR{\gamma}$) and CCAAT enhancer-binding-proteins ${\alpha}$ ($C/EBP{\alpha}$). This study potentially suggests that the Saekso 1 corn husk and cob extracts may improve lipid metabolism and reduce lipid accumulation.

Comparisons of Characteristics of Amaranth Starches Isolated from five Cultivars Grown in Korea (국내산 아마란스로부터 분리한 전분의 특성 비교)

  • Choi, Cha-Ran;Choi, Hun-Jae;Kim, Sung-Ran;Lee, Jae-Hak;Shin, Mal-Shick
    • Korean Journal of Food Science and Technology
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    • v.32 no.2
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    • pp.252-257
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    • 2000
  • The physicochemical and pasting properties of amaranth starches isolated from five cultivars, Andy, Suvarna, Nu World, K266-1, K432, grown in Korea, were investigated. The shapes of starch granules were all polygonal and size was in the range $1.14{\sim}1.48\;{\mu}m$ for all five cultivars. X-ray diffractograms were shown typical A type diffraction patterns for all amaranth starches. The protein and crude lipid contents of starches were $0.13{\sim}0.23%$ and $0.01{\sim}0.05%$, respectively. The apparent amylose contents ranged from $2.79{\sim}4.35%$ and the water binding capacities were $128.05{\sim}135.80%$. The transmittances of 0.1% amaranth starch suspensions except K266-1 increased rapidly above $65^{\circ}C$, thereafter increased slowely. The initial pasting and peak temperature ranges of five cultivars by RVA were $71.3{\sim}73.7^{\circ}C$ and $81.5{\sim}84.0^{\circ}C$, respectively. The peak and cooling viscosities followed the order : Nu World>Andy>Suvarna>K432>>K266-1. Nu World was shown the highest peak (166), final (103) and cooling viscosities (30 RVU). K266-I exhibited the lowest setback (-38) of all five cultivars. Gelatinization (To) and peak temperature (Tp) of amaranth starches in DSC thermograms were $65.7{\sim}68.0^{\circ}C$ and $70.6{\sim}75.8^{\circ}C$, respectively. Enthalpies followed the order: K266-1

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