• BMB Reports
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• 제28권2호
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• pp.162-169
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• 1995

Rat brain succinic semialdehyde dehydrogenase (EC 1.2.1.24 SSADH) activity was detected in mitochondrial, cytosolic and microsomal fractions. Brain mitochondrial soluble SSADH was purified by ammonium sulfate precipitation, DEAE Sephacel, and 5'-AMP Sepharose 4B affinity chromatography. The purified enzyme was shown to consist of four identical subunits, and the molecular weight of a subunit was 55 kD. The $K_m$ for short chain aliphatic aldehydes and aromatic aldehydes were at the $10^{-3}M$ level but that for succinic semialdehyde was 2.2 ${\mu}M$. Either $NAD^+$ or $NADP^+$ can be used as a cofactor but the affinity for $NAD^+$ was 10 times higher than that for $NADP^+$. The brain cytosolic SSADH was also purified by ammonium sulfate precipitation, DEAE Sephacel, Blue Sepharose CL-6B and 5'-AMP Sepharose 4B affinity chromatography and its Km for short chain aliphatic aldehydes was at the $10^{-3}$ level but that for succinic semialdehyde was 3.3 ${\mu}M$. $NAD^+$ can be used as a cofactor for this enzyme. We suppose that both enzyme might participate in the oxidation of succinic semialdehyde, which is produced during GABA metabolism. The activity of both cytosolic and mitochondrial SSADH was markedly inhibited when the concentration of succinic semialdehyde was high. The reciprocal plot pattern of product inhibition and initial velocity indicated a sequential ordered mechanism for mitochondrial matrix SSADH. Chemical modification data suggested that amino acid residues such as cysteine, serine and lysine might participate in the SSADH reaction.

• Genomics & Informatics
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• 제13권2호
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• pp.45-52
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• 2015

Acute myeloid leukemia is a well characterized blood cancer in which the unnatural growth of immature white blood cell takes place, where several genes transcription is regulated by the micro RNAs (miRNAs). Argonaute (AGO) protein is a protein family that binds to the miRNAs and mRNA complex where a strong binding affinity is crucial for its RNA silencing function. By understanding pattern recognition between the miRNAs-mRNA complex and its binding affinity with AGO protein, one can decipher the regulation of a particular gene and develop suitable siRNA for the same in disease condition. In the current work, HOXA9 gene has been selected from literature, whose deregulation is well-established in acute myeloid leukemia. Four miRNAs (mir-145, mir-126, let-7a, and mir-196b) have been selected to target mRNA of HOXA9 (NCBI accession No. NM_152739.3). The binding interaction between mRNAs and mRNA of HOXA9 gene was studied computationally. From result, it was observed mir-145 has highest affinity for HOXA9 gene. Furthermore, the interaction between miRNAs-mRNA duplex of all chosen miRNAs are docked with AGO protein (PDB ID: 3F73, chain A) to study their interaction at molecular level through an in silico approach. The residual interaction and hydrogen bonding are inspected in Discovery Studio 3.5 suites. The current investigation throws light on understanding of AGO-assisted miRNA based gene silencing mechanism in HOXA9 gene associated in acute myeloid leukemia computationally.

• 한국동물위생학회지
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• 제44권4호
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• pp.185-194
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• 2021

Polyglutamine extension in the coding sequence of mutant huntingtin causes neuronal degeneration associated with the formation of insoluble polyglutamine aggregates in Huntington's disease (HD). Mutant huntingtin can form aggregates within the nucleus and processes of neurons possibly due to misfolding of the proteins. To better understand the mechanism by which an elongated polyglutamine causes aggregates, we have developed an in vitro binding assay system of polyglutamine tract from truncated huntingtin. We made GST-HD exon1 fusion proteins which have expanded polyglutamine epitopes (e.g., 17, 23, 32, 46, 60, 78, 81, and 94 CAG repeats). In the present emergence of new study adjusted nanotechnology on protein chip such as surface plasmon resonance strategy which used to determine the substance which protein binds in drug discovery platform is worth to understand better neurodegenerative diseases (i.e., Alzheimer disease, Parkinson disease and Huntington disease) and its pathogenesis along with development of therapeutic measures. Hence, we used strengths of surface plasmon resonance (SPR) technology which is enabled to examine binding specificity and explore targeted molecular epitope using its electron charged wave pattern in HD pathogenesis utilize conjugated mutant epitope of HD protein and its interaction whether wild type GST-HD interacts with mutant GST-HD with maximum binding affinity at pH 6.85. We found that the maximum binding affinity of GST-HD17 with GST-HD81 was higher than the binding affinities of GST-HD17 with other mutant GST-HD constructs. Furthermore, our finding illustrated that the mutant form of GST-HD60 showed a stronger binding to GST-HD23 or GST-HD17 than GST-HD60 or GST-HD81. These results indicate that the binding affinity of mutant huntingtin does not correlate with the length of polyglutamine. It suggests that the aggregation of an expanded polyglutamine might have easily occurred in the presence of wild type form of huntingtin.

• Molecules and Cells
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• 제24권1호
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• pp.119-124
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• 2007

TLR4 together with CD14 and MD-2 forms a pattern recognition receptor that plays an initiating role in the innate immune response to Gram-negative bacteria. Here, we employed the surface plasmon resonance technique to investigate the kinetics of binding of LPS to recombinant CD14, MD-2 and TLR4 proteins produced in insect cells. The dissociation constants ($K_D$) of LPS for immobilized CD14 and MD-2 were $8.7{\mu}m$, and $2.3{\mu}m$, respectively. The association rate constant ($K_{on}$) of LPS for MD-2 was $5.61{\times}10^3M^{-1}S^{-1}$, and the dissociation rate constant ($K_{off}$) was $1.28{\times}10^2S^{-1}$, revealing slow association and fast dissociation with an affinity constant $K_D$ of $2.33{\times}10^6M$ at $25^{\circ}C$. These affinities are consistent with the current view that CD14 conveys LPS to the TLR4/MD-2 complex.

• 대한건축학회논문집:계획계
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• 제35권1호
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• pp.83-92
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• 2019

Advertising signboards are designed for the visibility to affects recall and recognition of costumers. It is well know that the visibility from images is created by the value difference among colors. The research defines whether the background color combination of outdoor signboards is configured to maximize visibility, by a series of color value variation in complex commercial facilities. The subject of study is to examine how the visibility is made by the color combination since visibility cannot be obtained independently. Two steps of analysis were performed to confirm that the color composition of signboards was based on the color value difference. The first is to analyze that the entire colors of signboards are clearly categorized as different value groups. All components of colors, hue, value and chroma had been analyzed by color aesthetic measures to prove that the value variation has the only regularity and the principle of composition. The second step is a further verification with an ample amount of samples to determine whether series of signboards create a value altering pattern. The data for analysis is gained by colorimetric survey and the color data are used for exponentializing the degree of combining, which shows selective affinity between each pair colors.

• 생명과학회지
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• 제17권10호
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• pp.1347-1353
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• 2007

성장과정 중 흰쥐 신장에서 나타나는 복합당질의 변화를 알아보기 위해 18일 태자부터 성체에 이르는 신장을 형태적 관찰과 더불어 9가지 lectin (SBA, DBA, PNA, BSL-1, RCA-1, sWGA, UEA-1, LCA 및 Con A)으로 검색하였다. 신장 발생단계에서 성숙한 신원구조와 함께 미성숙한 구조물 즉 소포와 요관아 등이 생후 14일에 이르기까지 관찰되었으며 생후 21일에 이르러 성체와 유사한 구조적 특성을 보였다. 복합당질의 변화를 보면 사구체에서 RCA-1, LCA및 Con A에 반응을 나타내며 RCA-1 및 LCA는 태자와 신생쥐에서 일시적으로 증가하다 성체에서 관찰되지 않으나 Con A는 성장과 더불어 증가하였다. 근위곡요세관은 UEA-1을 제외한 모든 lectin에 반응하며 DBA, SBA, PNA, BSL-1, RCA-1 및 Con A반응이 성장과 더불어 증가하며 특히 RCA-1과 BSL-1반응이 현저하였다. 이에 비해 sWGA와 LCA반응은 성장과정에 일시적으로 증가하며 성체에 이를수록 감소하였다. 원위곡요세관도 근위곡요세관 유사하게 DBA, SBA, PNA, BSL-1 및 RCA-1반응은 성숙과 함께 증가하나 LCA반응은 성숙과정에 일시적으로 증가하며 성체에서 감소하였다. 집합관에서는 DBA, SBA, PNA, sWGA반응이 성숙과 동시에 증가하나 BSL-1, RCA-1, LCA반응은 미성숙관에서 일시적으로 증가하였다. 이상의 반응으로 보아 신장발생과정에서 형태적 기능적 성숙과 함께 다양한 복합당질의 변화를 보이는데 대체로 성숙에 따라 반응이 증가하는 복합당질군과 미성숙기에 일시적으로 증가하며 성체에서 감소하는 복합당질군으로 대별할 수 있었다. 이러한 출생전후 복합당질의 변화는 신장의 기능적 성숙과정과 연관성을 가지며 발생과정에서 현저한 변화를 나타내는 복합당질은 정상 신장발생에 대한 표지인자로 유용할 것이다.

• 한국수산과학회지
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• 제33권1호
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• pp.55-59
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• 2000

무지개 송어의 뇌하수체 및 배양액에 존재하는 GTH II 농도를 측정하기 위해 Avidin- Biotin complex를 이용한 sandwich EIA 계을 개발했다. Protein A sepharose affinity chromatography을 통해서 얻어진 연어 GTH II의 rabbit IgG에 biotinylation시킨 것 (Biotin-salmon GTH II rabbit IgG)을 제2 항체로 사용하였고, Non-Biotin salmon GTH II rabbit IgG는 단지 protein A sepharose affinity chromatography에서 얻어진 IgG를 제 1 항체로 사용하였다. EIA는 sandwich법에 의해서 이루어졌으며, 효소반응 기질로는 TMB(3,3'5,5-tetramethylbenzidine)를 이용했으며, 반응후 450 nm의 흡광도에서 automatic microplate reader로 측정하였다. 그 결과, $0.12\;{\~}\;125\;ng/ml$의 범위에서 용량반응곡선을 얻었으며, 측정감도 (최소 검출량)는 거의 0.58 ng/ml 정도 였다. 그리고 뇌하수체 추출물 및 배양액 각각의 희석곡선은 GTH II 표존곡선과 일치 하였다. 또한 이러한 GTH II의 표준곡선는 뇌하수체내 다른 peptide hormone와는 교차반응을 거의 나타내지 않았다. Testosterone을 처리한 미성숙 무지개 송어의 뇌하수체 세포배양계를 이용하여 sGnRH에 의한 GTH II 분비량을 본 sandwich EIA계와 RIA계를 비교 조사한 결과, 거의 같은 분비량을 나타냈을 뿐만아니라 같은 분비 pattern을 나타냈다. 이러한 결과로부터 본 sandwich법 EIA계에 의해서 연어과 어류의 뇌하수체 추출물 및 뇌하수체 배양액 중의 GTH II 함량 및 분비량을 측정하는데 있어서 안정된 assay계라고 생각되어진다.

• Restorative Dentistry and Endodontics
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• 제20권1호
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• pp.152-163
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• 1995

This experiment was performed to observe the adhesion pattern and microleakage in the gingival margin according to variation in the resin cement thickness which results from thickness of Die spacer. which is considered to effect the adaptability of the composite resin inlays. Clearfil CR inlays were fabricated on stone models with CR Sep applicated once and Nice fit twice, 4 times, and 6 times each. After 2nd curing within the CRC-100 oven, CR inlays were cemented with CR inlay cement. Dye(2% methylene blue) penetration and adhesion pattern were evaluated after sectioning of gingival margin into :3 pieces. The results were as follows ; 1. The thickness of resin cement showed unevenchanging pattern with that of die spacer, namely, it was increased until 4 times' application of Nice-Fit but was decreased with 6 times' application of that. 2. The degree of dye penetration wasn't affected by cement thickness within a limited value. 3. Most of dye penetration was shown through the interface between cement and enamel rather than the interface between cement and CR inlay. This shows that the affinity of resin cement for CR inlay was superior to the adhesive strength with tooth structure. 4. No gap was found at the interface between enamel and cement but some showed separation between dentin and cement. It is concidered that the contraction force of cement was less than the bond strength with the enamel. 5. Lots of voids were found in the CR inlay and resin cement. There was a pooling tendency of bonding agent and cement in the axiogingival line angle portion. 6. In some specimens, cracks were shown in enamel margin. From this it could be considered that cavity preparation and surface treatment weakened the tooth structure.

• 방사선산업학회지
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• 제5권1호
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• pp.47-54
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• 2011

In this study, we investigated to evaluate differential expression of genes encoding lipid transfer proteins (LTP) by acute and chronic gamma irradiation in rice. After acute and chronic gamma irradiation by 100 Gy and 400 Gy to rice plant, necrotic lesion was observed in the leaf blade and anthocyanin contents were increased. We isolated a total of 21 rice lipid transfer protein (LTP) genes in the TIGR database, and these genes were divided into four different groups on the basis of nucleotide sequences. The LTP genes also were classified as different four classes according to expression pattern using RT-PCR. Group A, B contained genes with increased expression and decreased expression in acute and chronic, respectively. Group C contained genes with contrasted expression pattern. Group D wasn't a regular pattern. But the specific affinity was not obtained between two grouping.

• Biotechnology and Bioprocess Engineering:BBE
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• 제9권2호
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• pp.118-126
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• 2004

Recombinant E.coli ACV 1003 (recA::lacZ) releasing ${\beta}$-galactosidase by a SOS regulon system, when exposed to DNA-damaging compounds, have been used to effectively monitor endocrine disruptors. Low enzyme activity of less than 10 units/mL, corresponding to a $\mu\textrm{g}$/L(ppb) range of an endocrine disruptor (tributyl tin, bisphenol A. etc.), can be rapidly determined, not by a conventional time-consuming and tedious enzyme assay, but by an alternative interferometric biosensor. Heavily boron-doped porous silicon for application as an interferometer, was fabricated by etching to form a Fabry-Perot fringe pattern, which caused a change in the refractive index of the medium including ${\beta}$-galactosidase. In order to enhance the immobilization of the porous silicon surface, a calyx crown derivative (ProLinker A) was applied, instead of a conventional biomolecular affinity method using biotin. This resulted in a denser linked formation. The change in the effective optical thickness versus ${\beta}$-galactosidase activity, showed a linear increase up to a concentration of 150 unit ${\beta}$-galactosidase/mL, unlike the sigmoidal increase pattern observed with the biotin.