• Fisheries and Aquatic Sciences
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• v.1 no.2
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• pp.201-208
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• 1998

A protease, which had no tryptic and chymotryptic activity, was purified from the hepatopancreas of shrimp, P. japonicus, through ammonium sulfate fractionation, Q­Sepharose ionic exchange, benzamidine Sepharose 6B affinity, and Sephacryl S-100 gel chromatography. Molecular weight (M.W.) of the protease was estimated to be 24 kDa by gel filtration and showed a single peptide band by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE). The protease had a low ratio of acidic to basic amino acids, which is different with pro teases from marine animals. The enzyme was partially inhibited by benzamidine, tosyl-L-lysine chioromethyl ketone (TLCK), phenylmethylsulfonyl fluoride (PMSF), soybean trypsin inhibitor (SBTI), and pepstatin. The enzyme did not have any activity against benzoyl-D,L-arginine p-nitroanilide (BAPNA) or benzoyl-L-tyrosine ethyl ester (BTEE) which is a specific substrate of trypsin and chymotrypsin, respectively. However, the enzyme showed activity forward N-CBZ-L-tyrosine p-nitrophenyl ester (CBZ-Tyr-pNE), N­CBZ-L-tryptophan p-nitrophenyl ester (CBZ-Trp-pNE), and N-CBZ-L-proline p-nitrophenyl ester (CBZ-Pro-pNE). The protease did not showed tryptic and chymotryptic activity, which was not reported in shrimp hepatopancreas.

• Journal of the Korean Society of Food Science and Nutrition
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• v.27 no.4
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• pp.745-750
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• 1998

The antimutagenic and antigenotoxic effects of ethanol, methanol, water and non-heating ethanol extract of Ligularia fischeri were investigated using Ames test and micronucleus test. Four solvent extracts by themseleves did not induce mutagenesis. The four extract of 200㎍/plate showed approximately 84.7%, 77.1%, 72.5% and 71% inhibitory effect on the mutagenesis induced by N-methyl-N'-nitro-N-nitrosoguanidine(MNNG) and 67.9%, 66.8%, 64.6% and 56% inhibition on the mutagenesis by 4-nitroquinoline-1-oxide(4NQO) against TA100 strain, whereas 70.2%, 60.9%, 61.9% and 52.8% inhibitions were observed on the mutagenesis induced by 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indol(Trp-P-1) in the presence of 200㎍/plate. TA100 strain was more sensitive than TA98 strain by four kinds of extracts on antimutagenesis. The effects of Ligularia fischeri extracts on the frequencies of micronucleated poly chromatic erythrocytes(MNPECs) induced by MNNG were investigated in the bone marrow. Ten, 20, 40 and 80mg g/kg of each extract were administered to animals immediately after injection of MNNG and the exposure time was 36 hours. Inhibitory effects of Ligularia fischeri ethanol extracts were 12%, 35.3%, 58.8%, and 57%, in the presence of 20, 40, 60 and 80mg/kg, respectively whereas methanol extracts showed 15.5%, 32.7%, 50.8%, and 57.9% inhibitory effects, respectively. Both extracts showed enhanced antimutagenic and antigenotoxic effects. These results showed a good correlation between antimutagenic effects in in vitro and in in vitro assay.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.2
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• pp.173-177
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• 2000

연구목적: 한국여성의 다낭성 난포증후군 환자에서 황체형성 호르몬 exon 2 유전자의 변이를 탐색하여 이들 변이와 질환과의 관련성 여부를 밝히고자 하였다. 연구재료 및 방법: 21명의 다낭성 난포증후군 환자를 대상으로 황체형성 호르몬 exon 2(Trp8Arg;TGG to CGG and Ile15Thr; ATC to ACC)의 변이를 탐색하였다. 혈액에서 Genomic DNA를 추출하여 PCR로 증폭한 후 RFLP 방법으로 변이형을 구분하였다. 결과: 황체형성 호르몬 exon 2의 변이형이 다낭성 난포증후군 환자에서 28.6%로 이미 조사된 바 있는 대조군의 16.7% 보다 약간 높게 나타났으나 통계적으로 유의한 차이는 없었다(p>0.05). 결론: 황체형성 호르몬 exon 2의 변이가 한국인의 다낭성 난포증후군 발병과 관련이 있는지를 밝히기 위해 더 많은 개체에 대한 연구가 요구된다.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.2
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• pp.179-182
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• 2000

This study was performed to determine whether the $LH{\beta}$-subunit gene missense mutation is present in Korean infertile patients with 46,XX POF women. The variants of $LH{\beta}$ exon 2 (Trp 8Arg; TGG to CGG and Ile15Thr; ATC to ACC) were studied in forty-four 46,XX idiopathic POF and 54 nonpregnant women. The $LH{\beta}$ exon 2 variants were more frequent in POF patients (20.5%) than nonpregnant (16.7%) women (p>0.05). POF patients with the variant was slightly higher than nonpregnant women with the variant.

• Bulletin of the Korean Chemical Society
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• v.32 no.10
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• pp.3675-3681
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• 2011

Fusarium oxysporum, an important pathogen that mainly causes vascular or fusarium wilt disease which leads to economic loss. Disruption of gene encoding a heterotrimeric G-protein-${\beta}$-subunit (FGB1), led to decreased intracellular cAMP levels, reduced pathogenicity, colony morphology, and germination. The plant defense protein, Nicotiana alata defensin (NaD1) displays potent antifungal activity against a variety of agronomically important filamentous fungi. In this paper, we performed a molecular modeling and docking studies to find vital amino acids which can interact with various antifungal compounds using Discovery Studio v2.5 and GRAMMX, respectively. The docking results from FGB1-NaD1 and FGB1-antifungal complexes, revealed the vital amino acids such as His64, Trp65, Ser194, Leu195, Gln237, Phe238, Val324 and Asn326, and suggested that the anidulafungin is a the good antifungal compound.The predicted interaction can greatly assist in understanding structural insights for studying the pathogen and host-component interactions.

• Animal cells and systems
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• v.14 no.2
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• pp.99-114
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• 2010

With the consensus sequence information of the serum glucocorticoid-induced protein kinase-1 (SGK1) phosphorylation site {R-X-R-X-X-(S/T)$\Phi$; where $\Phi$ is any hydrophobic amino acid}, we noticed that the transient receptor potential vanilloid 4 (TRPV4) cation channel, a member of the TRP vanilloid subfamily, harbors the putative SGK1 phosphorylation site (on its Ser 824). We have demonstrated that TRPV4 is an SGK1 authentic substrate protein, with the phosphorylation on the Ser 824 of TRPV4 by SGK1. Further, using TRPV4 mutants (S824A and S824D), we noted that the modification of the Ser 824 activates its $Ca^{2+}$ entry, and sensitizes the TRPV4 channel to 4-$\alpha$-phorbol 12,13-didecanoate (4-${\alpha}PDD$) or heat, simultaneously enhancing its active state. Additionally, we determined that the modification of the Ser 824 controls both its plasma membrane localization and its protein interactions with calmodulin. Thus, we have proposed herein that phosphorylation on the Ser 824 of TRPV4 is one of the control points for the regulation of its functions.

• Bulletin of the Korean Chemical Society
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• v.28 no.6
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• pp.947-952
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• 2007

Mycobacterium tuberculosis is a pathogen responsible for 2-3 million deaths every year worldwide. The emergence of drug-resistant and multidrug-resistant tuberculosis has increased the need to identify new antituberculosis targets. Acetohydroxy acid synthase, (AHAS, EC 2.2.1.6), an enzyme involved in branched-chain amino acid synthesis, has recently been identified as a potential anti-tuberculosis target. To assist in the search for new inhibitors and “receptor-based” design of effective inhibitors of tubercular AHAS (TbAHAS), we constructed four different structural models of TbAHAS and used one of the models as a target for virtual screening of potential inhibitors. The quality of each model was assessed stereochemically by PROCHECK and found to be reliable. Up to 89% of the amino acid residues in the structural models were located in the most favored regions of the Ramachandran plot, which indicates that the conformation of each residue in the models is good. In the models, residues at the herbicide-binding site were highly conserved across 39 AHAS sequences. The binding mode of TbAHAS with a sulfonylurea herbicide was characterized by 32 hydrophobic interactions, the majority of which were contributed by residue Trp516. The model based on the highest resolution X-ray structure of yeast AHAS was used as the target for virtual screening of a chemical database containing 8300 molecules with a heterocyclic ring. We developed a short list of molecules that were predicted to bind with high scores to TbAHAS in a conformation similar to that of sulfonylurea derivatives. Five sulfonylurea herbicides that were calculated to efficiently bind TbAHAS were experimentally verified and found to inhibit enzyme activity at micromolar concentrations. The data suggest that this time-saving and costeffective computational approach can be used to discover new TbAHAS inhibitors. The list of chemicals studied in this work is supplied to facilitate independent experimental verification of the computational approach.

• Korean Journal of Food Science and Technology
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• v.35 no.2
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• pp.325-328
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• 2003

Nine commercial soy sauces $(A{\sim}I)$ were investigated for their biogenic amine (BAs) levels. Detected biogenic amines were putrescine (PUT), cadaverine (CAD), trytamine (TRP), ${\beta}-phenylethylamine$ (PHE), spermine (SPM), histamne (HIS), and tyramine (TYR). All products tested had biogenic amines as detected level. PUT was the major biogenic amines detected in six products, and difference between the highest and the lowest among products was more than 16 mg/kg. Six products had all seven biogenic amines tested, while one product had only five. Results indicate that soy sauces commercially available in Korea contain biogenic amines at various levels. Studies related to biogenic amines including survey of contents must be performed continuously.

• The Korean Journal of Mycology
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• v.22 no.3
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• pp.247-253
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• 1994

This study was carried out to isolate and characterize $A{\alpha}$ mating locus controlling fruiting body formation directly in the Basidiomycete Schzophyllum commune growing in the North America. Total numbers of genomic library of S. commune UVM1-34 was about $2{\times}10^4$ cells. About 90% library was appeared to have about 35 kb inserted genome DNA in cosmid pTC20 vector. 6 clones were proved to have positive signal to probes within Z and Y region in colony and southern hybridization. In the mating activity test, all the 6 positive clones were appeared to have $A{\alpha}3$ mating activity although they had two different restriction patterns. pSC13 containing 5.7 Kb PstI-fragment of UVM 1-34 $A{\alpha}3$ allele showed about 50% clamp cell formation indicating mating activity when cotransformation was done together with cosmid pTC20.

• Journal of Microbiology and Biotechnology
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• v.22 no.12
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• pp.1724-1730
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• 2012

An ${\alpha}$-L-arabinofuranosidase (TmAFase) from Thermotoga maritima MSB8 is a highly thermostable exo-acting hemicellulase that exhibits a relatively higher activity towards arabinan and arabinoxylan, compared with other glycoside hydrolase 51 family enzymes. In the present study, we carried out the enzymatic characterization and structural analysis of TmAFase. Tight domain associations found in TmAFase, such as an inter-domain disulfide bond (Cys306 and Cys476) in each monomer, a novel extended arm (amino acids 374-385) at the dimer interface, and total 12 salt bridges in the hexamer, may account for the thermostability of the enzyme. One of the xylan binding determinants (Trp96) was identified in the active site, and a region of amino acids (374-385) protrudes out forming an obvious wall at the substrate-binding groove to generate a cavity. The altered cavity shape with a strong negative electrostatic distribution is likely related to the unique substrate preference of TmAFase towards branched polymeric substrates.