• 대한약리학회지
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• 제30권2호
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• pp.255-260
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• 1994

There are some reports about the influences of free fatty acids on the albumin binding of drugs. But they were concerned to the limited free fatty acids, mostly of azapropazone-warfarin bidning site bound drugs and determination of dissociation and association constants by stopped flow technique. These data were not enough to make conculsions for the general tendency of free fatty acid to albumin binding. Therefore the influence of various saturated fatty acids of $C_{10{\sim}20}$, oleic acid and linoleic acid as unsaturated fatty acids to albumin binding of warfarin and dansylsarcosine were studied by equilibrium dialysis. The concentration of free drug was determined by spectrophotometer according to the molar ratios of 0, 0.5, 1, 2 and 4 between free fatty acid and albumin. There were significant increasing in the free durg concentration of warfarin and dansylsarcosine when the molar ratio for capric acid, lauric acid and palmitic acid was 4. The free warfarin concentration was increased significantly at a molar ratio of 4 between oleic acid and albumin. Therefore the albumin binding of durgs can be variated significantly by increased free fatty acid of diabetics and cause to the pharmacokinetic variation between healthy and diabetics.

• Weed & Turfgrass Science
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• 제6권1호
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• pp.28-31
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• 2017

The mutation rate of proline in the position 197 (Pro197) in acetohydroxy acid synthase (AHAS) is highest among sulfonylurea (SU) herbicide-resistance mutants. Therefore, it is significant to investigate the resistance mechanism for the mutation and to develop the herbicides specific to the mutants. SU herbicide resistance mechanism of the mutants, 197Ser, 197Thr and 197Ala, in AHAS were targeted for designing new SU-herbicide. We did molecular dynamics (MD) simulation for understanding SU herbicide-resistance mechanisms of AHAS mutants and designed new herbicides with docking and MD evaluations. We have found that mutation to 197Ala and 197Ser enlarged the entrance of the active site, while 197Thr contracted. Map of the root mean square derivation (RMSD) and radius gyrations (Rg) revealed the domain indicating the conformations for herbicide resistant. Based on the enlarging-contracting mechanism of active site entrance, we designed new herbicides with substitution at the heterocyclic moiety of a SU herbicide for the complementary binding to the changed active site entrances of mutants, and designed new herbicides. We confirmed that our screened new herbicides bonded to both AHAS wild type and mutants with higher affinity, showing more stable binding conformation than the existing herbicides.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2001년도 추계학술대회 및 정기총회
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• pp.94-94
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• 2001

Prostaglandin endoperoxide H synthase (PGHS) catalyzes the committed step in prostaglandins and thromboxane A$_2$-- oxygenation of arachidonic acid to the hydroperoxy endoperoxide PGG$_2$, followed by reduction PGG$_2$to the alcohol PGH$_2$. The two reactions by PGHS -- cyclooxygenase and peroxidase -- occur at distinct but structurally and functionally interconnected sites. The peroxidase reaction occurs at a heme-containing active site located near the protein surface. The cyclooxygenase reaction occurs in a hydrophobic channel in the core of the enzyme. Initially a peroxide reacts with the heme group, yielding Compound I and an alcohol derived from the oxidizing peroxide. Compound I next undergoes an intramolecular reduction by a single electron traveling from Tyr385 along the peptide chain to the proximal heme ligand, His388, and finally to the heme group. Following the binding of arachidonic acid, Tyr385 tyrosyl radical initiates the cyclooxygenase reaction by abstracting the 13-pro(5) hydrogen atom to give an arachidonyl radical, which sequentially reacts with two molecules of oxygen to yield PGG$_2$. In order to characterize PGHS peroxidase active site, we examined various lipid peroxides with purified recombinant ovine PGHS proteins and determined the rate constants. The results have shown that twenty-carbon unsaturated fatty acid hydroperoxides have similar efficiency in peroxidation by PGHS, irrespective of either the location of hydroperoxy group or the number of double bonds. It was also confirmed by the subsequent study with PGHS peroxidase active site mutants.

• 대한화학회지
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• 제23권3호
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• pp.161-164
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• 1979

$Ca^{2+},\;La^{3+},\;NH_4^+$ 등으로 양이온을 교혼하거나, 알루미늄을 추출한 합성 zeolite Y에 팔라듐을 담지시켜, 촉매산성도 및 담지된 금속의 유효표면적을, n-butane의 분해반응에서 촉매활성과 관련지어 조사하였다. 암모니아의 TPD(Temperature Programmed Desorption) 실험에 의하면 NaY < CaY < LaY 순으로 강한 산점의 양이 많아졌으며, 알루미늄 추출로 $SiO_2/Al_2O_3$의 비가 커질 수록 전체산점의 양은 감소되었으나, 강한 산점의 양은 증가되었다. CO의 화학흡착으로 측정한 담지금속의 유효표면적은 산성도가 큰 촉매에서 비교적 컸다. n-Butane의 zeolite 촉매에서의 반응은, 촉매의 산성도와 금속성분의 유효표면적에 관계되나, 전화율이 측정한 범위내에서의 유효표면적에 비례하므로 금속성분에 의한 탈수소반응이 중요한 단계로 생각된다.

• 대한화학회지
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• 제20권6호
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• pp.441-447
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• 1976

여러가지 양이온이 교환된 silicate 상에 pyridine, tertiary butylamine, ethylenediamine을 흡착시켜 IR spectra를 4000 ∼ 1200 $cm^{-1}$의 범위에서 그리고 여러 다른 탈기온도에서 얻었다. 이 결과 protonic 산 자리와 aprotonic 산 자리를 구별할 수 있었으며 양이온이 교환된 모든 silicate는 Bronsted 산과 Lewis 산을 모두 나타내었다. 그리고$ Na^+$이온이 교환된 silicate가 adsorbate와의 반응성이 가장 적게 나타났다. Tertiary butylamine의 band intensity의 상대적인 비는 교환된 양이온의 polarizing power와 비례하고 두개의 amino group을 가지고 있는 ethylenediamine은 tertiary butylamine에 비해서 silicate 표면으로부터 쉽게 탈착되지 않았으며 또한 Lewis 산 자리와 coordination bond를 이루는 경우와 Bronsted 산 자리와 $NH3^+$ 종을 만드는 경우외에 migrating proton을 떼어 냄으로 말미암아 표면산소와의 수소결합을 이루는 경우가 조사되었다.

• Journal of Microbiology and Biotechnology
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• 제16권9호
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• pp.1434-1440
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• 2006

The stable anchorage of pyridoxal 5'-phosphate (PLP) in the active site of D-amino acid transaminase (D-AT) is crucial for the enzyme catalysis. The three-dimensional structure of D-AT revealed that Glu32 is one of the active site groups that may playa role in PLP binding. To prove the role of Glu32 in PLP stability, we firstly checked the rate of the potential rate-limiting step. The kinetic analysis showed that the rate of the ${\alpha}$-deprotonation step reduced to 26-folds in E32A mutant enzyme. Spectral analyses of the reaction of D-AT with D-serine revealed that the E32A mutant enzyme failed to stabilize the key enzyme-substrate intermediate, namely a quinonoid intermediate (E-Q). Finally, analysis of circular dichroism (CD) on the wild-type and E32A mutant enzymes showed that the optical activity of PLP in the enzyme active site was lost by the removal of the carboxylic group, proving that Glu32 is indeed involved in the cofactor anchorage. The results suggested that the electrostatic interaction network through the groups from PLP, Glu32, His47, and Arg50, which was observed from the three-dimensional structure of the enzyme, plays a crucial role in the stable anchorage of the cofactor to give necessary torsion to the plane of the cofactor-substrate complex.

• Food Science and Biotechnology
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• 제17권6호
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• pp.1221-1227
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• 2008

The application of the cellulase gene (celA) as a selection marker of food-grade integration system was investigated in Lactobacillus (Lb.) casei, Lactococcus lactis, and Leuconostoc (Leu.) mesenteroides. The 6.0-kb vector pOC13 containing celA from Clostridium thermocellum with an integrase gene and a phage attachment site originating from bacteriophage A2 was used for site-specific recombination into chromosomal DNA of lactic acid bacteria (LAB). pOC13 was also equipped with a broad host range plus replication origin from the lactococcal plasmid pWV01, and a controllable promoter of nisA ($P_{nisA}$) for the production of foreign proteins. pOC13 was integrated successfully into Lb. casei EM116, and pOC13 integrants were easily detectable by the formation of halo zone on plates containing cellulose. Recombinant Lb. casei EM 116::pOC13 maintained these traits in the absence of selection pressure during 100 generations. pOC13 was integrated into the chromosome of L. lactis and Leu. mesenteroides, and celA acted as an efficient selection marker. These results show that celA can be used as a food-grade selection marker, and that the new integrative vector could be used for the production of foreign proteins in LAB.

• Bulletin of the Korean Chemical Society
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• 제28권5호
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• pp.772-776
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• 2007

In order to study the role of residue in the active site of glutathione S-transferase (GST), Arg13 residue in human GST P1-1 was replaced with alanine, lysine and leucine by site-directed mutagenesis to obtain mutants R13A, R13K and R13L. These three mutant enzymes were expressed in Escherichia coli and purified to electrophoretic homogeneity by affinity chromatography on immobilized GSH. Mutation of Arg13 into Ala caused a substantial reduction of the specific activity by 10-fold. Km GSH, Km DCNB and Km EPNP values of R13A were approximately 2-3 fold larger than those of the wild type. Mutation of Arg13 into Ala also significantly affected I50 values of S-methyl-GSH that compete with GSH and ethacrynic acid, an electrophilic substrate-like compound. These results appeared that the substitution of Arg13 with Ala resulted in significant structural change of the active site. Mutation of Arg13 into Leu reduced the catalytic activity by approximately 2-fold, whereas substitution by Lys scarcely affected the activity, indicating the significance of a positively charged residue at position 13. Therefore, arginine 13 participates in catalytic activity as mainly involved in the construction of the proper electrostatic field and conformation of the active site in human GST P1-1.

• The Korean Journal of Physiology and Pharmacology
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• 제2권3호
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• pp.385-393
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• 1998

Human neutrophil elastase (HNElastase, EC 3.4.21.37), a causative factor of inflammatory diseases, was purified by Ultrogel AcA54 gel filtration and CM-Sephadex ion exchange chromatography. HNElastase was inhibited by phenylbutazone in a concentration dependent manner up to 0.4 mM, but as the concentration increased, the inhibitory effect gradually diminished. Binding of phenylbutazone to the human neutrophil elastase caused strong Raman shifts at 200, 440, and 1194 $cm^{-1}$. The peak at 1194 $cm^{-1}$ might be evidence of the presence $of\;-N=N-{\Phi}$ radical. The core area of the elastase, according to the visual molecular model of human neutrophil elastase, was structurally stable. A deeply situated active center was at the core area surrounded by hydrophobic amino acids. Directly neighboring the active site was one positively charged atom and two atoms carrying a negative charge, which enabled the enzyme and the drug to form a strong interaction. Phenylbutazone may form a binding, similar to a key & lock system to the atoms carrying opposite charges near the active site of the enzyme molecule. Furthermore, the hydrophobicity of the surrounding amino acid near the active site seemed to enhance the binding strength of phenylbutazone. Binding of phenylbutazone near the active site may cause masking of the active site, preventing the substrate from approaching the active site and inhibiting elastase activity.

• 대한바이러스학회지
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• 제28권4호
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• pp.303-316
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• 1998

An attenuated classical swine fever virus (CSFV), Suri strain, is a variant derived from a vaccine virus, LOM strain. This study was performed to elucidate the molecular biologcal properties of CSFV Suri strain, and to obtain the basic data for molecular epidemiological approaches for the disease. The truncated form of gp55 gene without the C-terminal transmembrane domain, in size of 1,023bp, was amplified by RT-PCR and sequenced by dye terminator cyclic sequencing method, and inserted into BamHI site of pAcGP67B baculovirus vector, establishing a cloned pAcHEG plasmid. By the nucleotide sequences determined, 341 amino acid sequences were predicted. As compared the nucleotide and amino acid sequences of gp55 of Suri with the various CSFV, Suri strain showed the high homology over 99.1% with ALD and LOM strains, but comparably the lower homology with Alfort and Brescia. In comparison of amino acid sequence in variable domain of gp55 protein, the similar tendency of homology was observed. In hydrophobicity analysis, all of four CSFV strains revealed the analogous patterns of hydrophobicity. The numbers and locations of N-glycosylation site and cysteine residues in gp55 were analyzed, those of Suri strain being coincident with ALD and LOM strains. The results suggest that gp55 in Suri strain has the high similarity to those in ALD and LOM strains in terms of the nucleotide and amino acid sequences and the functional properties of gp55 protein.