• Title/Summary/Keyword: S-247

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A Role for Leu247 Residue within Transmembrane Domain 2 in Ginsenoside-Mediated α7 Nicotinic Acetylcholine Receptor Regulation

  • Lee, Byung-Hwan;Choi, Sun-Hye;Pyo, Mi Kyung;Shin, Tae-Joon;Hwang, Sung-Hee;Kim, Bo-Ra;Lee, Sang-MoK;Lee, Jun-Ho;Lee, Joon-Hee;Lee, Hui Sun;Choe, Han;Han, Kyou-Hoon;Kim, Hyoung-Chun;Rhim, Hyewhon;Yong, Joon-Hwan;Nah, Seung-Yeol
    • Molecules and Cells
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    • v.27 no.5
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    • pp.591-599
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    • 2009
  • Nicotinic acetylcholine receptors (nAChRs) play important roles in nervous system functions and are involved in a variety of diseases. We previously demonstrated that ginsenosides, the active ingredients of Panax ginseng, inhibit subsets of nAChR channel currents, but not ${\alpha}7$, expressed in Xenopus laevis oocytes. Mutation of the highly conserved Leu247 to Thr247 in the transmembrane domain 2 (TM2) channel pore region of ${\alpha}7$ nAChR induces alterations in channel gating properties and converts ${\alpha}7$ nAChR antagonists into agonists. In the present study, we assessed how point mutations in the Leu247 residue leading to various amino acids affect 20(S)-ginsenoside $Rg_3$ ($Rg_3$) activity against the ${\alpha}7$ nAChR. Mutation of L247 to L247A, L247D, L247E, L247I, L247S, and L247T, but not L247K, rendered mutant receptors sensitive to $Rg_3$. We further characterized $Rg_3$ regulation of L247T receptors. We found that $Rg_3$ inhibition of mutant ${\alpha}7$ nAChR channel currents was reversible and concentration-dependent. $Rg_3$ inhibition was strongly voltage-dependent and noncompetitive manner. These results indicate that the interaction between $Rg_3$ and mutant receptors might differ from its interaction with the wild-type receptor. To identify differences in $Rg_3$ interactions between wild-type and L247T receptors, we utilized docked modeling. This modeling revealed that $Rg_3$ forms hydrogen bonds with amino acids, such as Ser240 of subunit I and Thr244 of subunit II and V at the channel pore, whereas $Rg_3$ localizes at the interface of the two wild-type receptor subunits. These results indicate that mutation of Leu247 to Thr247 induces conformational changes in the wild-type receptor and provides a binding pocket for $Rg_3$ at the channel pore.

Biochemical Characteristics for the Cofactor Free Mutant of Yeast Homocysteine Catalyzing Enzyme, Cystathionine ${\beta}$-Synthase (조효소를 함유하지 않는 효모의 Homocysteine 분해효소, Cystathionine ${\beta}$-Synthase의 생화학적 특성)

  • Jhee, Kwang-Hwan;Cho, Hyun-Nam;Yang, Seun-Ah;Lee, In-Seun
    • Microbiology and Biotechnology Letters
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    • v.35 no.3
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    • pp.196-202
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    • 2007
  • Mutations in the cystathionine ${\beta}$-synthase (CBS) gene cause homocystinuria, the most frequent inherited disorder in sulfur metabolism. CBS is the unique enzyme using both heme and pyridoxal 5-phosphate (PLP) for activity. Among the reported 140 mutations, one of the most common disease-causing alterations in human CBS is G307S mutation. To investigate the pathogenic mechanism of G307S by spectroscopic methods, we engineered the full length and the truncated G247S mutation of yeast CBS that is corresponding mutation to human G307S. Yeast CBS does not contain heme and thus gives a merit to study the spectroscopic properties. The UV-visible spectra of the purified full length and the truncated G247S yeast CBSs showed the total absence of PLP in the protein. The absence of PLP in G247S mutation was also confirmed by the PLP-cyanide adduct formation experiment, which was conducted by the incubation of the purified enzyme with KCN. The adducts were detected using a circular dichroism (CD) and a spectrofluorimeter. Radio isotope activity assay of full length and truncated G247S proteins also gave no activity. Our yeast G247S mutation data suggested that G307S might make the distortion of the active site so that cofactor PLP and substrate can not fit inside the active site. Our yeast CBS study addressed the reason why the G307S mutation in human CBS makes the enzyme inactive that consequently leads to severe clinical phenotype.

Estimation of Liquid Physical Properties of Mar-M247LC Superalloy by Directional Solidification (일방향 응고법을 이용한 Mar M-247LC 초내열합금의 액상 물성 측정)

  • Kim, Hyeon-Cheol;Lee, Jae-Hyeon;Seo, Seong-Mun;Kim, Du-Hyeon;Jo, Chang-Yong
    • Korean Journal of Materials Research
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    • v.11 no.9
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    • pp.721-726
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    • 2001
  • Directional solidification experiments have been carried out at the solidification rates from 0.5 to 50$\mu\textrm{m}$/s in Mar M-247LC superalloy in which several important liquid properties were estimated by analyzing the interface stability and temperature gradient at the solid/liquid interface. The diffusion coefficient in the liquid was estimated by employing the constitutional supercooling criterion. The temperature gradients changed with solidification rates and latent heat of solidification. The thermal conductivities of solid and liquid could be estimated by heat flux balance at the solid liquid interface.

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Isolation, Identification and Optimal Culture Conditions of Streptomyces albidoflavus C247 Producing Antifungal Agents against Rhizoctonia solani AG2-2

  • Islam, Rezuanul;Jeong, Yong-Tae;Ryu, Yeon-Ju;Song, Chi-Hyun;Lee, Yong-Se
    • Mycobiology
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    • v.37 no.2
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    • pp.114-120
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    • 2009
  • Streptomyces albidoflavus C247 was isolated from the soil of the Gyeongsan golf course in Korea. Physiological, biochemical and 16S rDNA gene sequence analysis strongly suggested that the isolate belonged to Streptomyces albidoflavus. Preliminary screening revealed that the isolate was active against fungi and bacteria. Self-directing optimization was employed to determine the best combination of parameters such as carbon and nitrogen source, pH and temperature. Nutritional and culture conditions for the production of antibiotics by this organism under shake-flask conditions were also optimized. Maltose (5%) and soytone (5%) were found to be the best carbon and nitrogen sources for the production of antibiotics by S. albidoflavus C247. Additionally, 62.89% mycelial growth inhibition was achieved when the organism was cultured at $30^{\circ}C$ and pH 6.5. Ethyl acetate (EtOAc) was the best extraction solvent for the isolation of the antibiotics, and 100 ${mu}$/ml of EtOAc extract was found to inhibit 60.27% of the mycelial growth of Rhizoctonia solani AG2-2(IV) when the poison plate diffusion method was conducted.

ERRATUM: Patient's Perception for Developing Research Questions of a Clinical Practice Guideline of Korean Medicine for Degenerative Lumbar Spinal Stenosis: A Descriptive Cross-Sectional Survey (ERRATUM: 퇴행성 요추 척추관 협착증의 한의표준임상진료지침 핵심질문 설정에 대한 환자 인식 설문 조사: 단면 설문 연구)

EXPLICIT MINIMUM POLYNOMIAL, EIGENVECTOR AND INVERSE FORMULA OF DOUBLY LESLIE MATRIX

  • WANICHARPICHAT, WIWAT
    • Journal of applied mathematics & informatics
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    • v.33 no.3_4
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    • pp.247-260
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    • 2015
  • The special form of Schur complement is extended to have a Schur's formula to obtains the explicit formula of determinant, inverse, and eigenvector formula of the doubly Leslie matrix which is the generalized forms of the Leslie matrix. It is also a generalized form of the doubly companion matrix, and the companion matrix, respectively. The doubly Leslie matrix is a nonderogatory matrix.