• 제목/요약/키워드: residue interaction analysis

검색결과 36건 처리시간 0.02초

환경 독성 Peptide의 인지질과의 상호 작용 특성 분석 (Analysis of the Interactive Characteristic of Environmental Toxic Peptide and Phospholipid)

  • 이봉헌;박흥재
    • 한국환경과학회지
    • /
    • 제12권1호
    • /
    • pp.77-80
    • /
    • 2003
  • The interaction of mastoparan B, a cationic tetradecapeptide amide isolated from the hornet Vespa basalis, with phospholipid bilayers was studied with synthetic mastoparan B and its analogue with Ala instead of hydrophobic 12th amino acid residue in mastoparan B. MP-B and its derivative, [12-Ala]MP-B were synthesized by the solid-phase peptide synthesis method. MP-B and its analogue, [12-Ala]MP-B adopted an unordered structure in buffer solution. In the presence of neutral and acidic liposomes, the peptides took an $\alpha$-helical structure. The two peptides interacted with neutral and acidic lipid bilayers. These results indicated that the hydrophobic face in the amphipathic $\alpha$-helix of MP-B critically affected the biological activity and helical content.

Effects of the Heptasequence SPTSPTY of Rat Nuclear Factor 1-A on Interactions between the C-Terminal Regions of Mammalian Nuclear Factor 1 Proteins

  • Hwang, Jung-Su;Kim, Ji-Young
    • BMB Reports
    • /
    • 제33권6호
    • /
    • pp.519-524
    • /
    • 2000
  • NF1 proteins are a family of DNA binding proteins which consist of two separate domains, N-terminal DNA binding domain and C-terminal transcription activation domain. The N-terminal 220 amino acids are highly conserved and are also known to mediate dimerization of NF1 proteins. The C-terminal regions of different type of NF1 proteins are heterogeneous and responsible for transcriptional activation. In this study, we tested the interaction between different domains of rat NF1-A protein by yeast two hybrid analysis and observed the interaction between C-terminal regions of NF1-A which do not contain the N-terminal dimerization domain. Our results showed that the C-terminal region of rat NF1-A between residues 231 and 509 strongly interacted not only with itself, but also with human NF1/CTF1 which is a different type of NF1. When the C-terminal region was divided into two fragments, one from residue 231 to 447 and the other from 448 to 509, the two fragments were able to interact with the C-terminal region of NF1-A significantly. This indicates that both fragments contain independent interaction domains. Analysis of the interactions with alanine substituted fragments showed that substitutions of the heptasequence, SPTSPTY of NF1-A, affected interaction between NF1 proteins. Our results strongly suggest that C-terminal regions may also be important for the formation of homo- and heterodimers in addition to the N-terminal dimerization domain. Also, the heptasequence motif may play some roles in dimer formation.

  • PDF

Folding Mechanism of WT* Ubiquitin Variant Studied by Stopped-flow Fluorescence Spectroscopy

  • Park, Soon-Ho
    • Bulletin of the Korean Chemical Society
    • /
    • 제31권10호
    • /
    • pp.2877-2883
    • /
    • 2010
  • The folding kinetics of $WT^*$ ubiquitin variant with valine to alanine mutation at sequence position 26 (HubWA) was studied by stopped-flow fluorescence spectroscopy. While unfolding kinetics showed a single exponential phase, refolding reaction showed three exponential phases. The semi-logarithmic plot of urea concentration vs. rate constant for the first phase showed v-shape pattern while the second phase showed v-shape with roll-over effect at low urea concentration. The rate constant and the amplitude of the third phase were constant throughout the urea concentrations, suggesting that this phase represents parallel process due to the configurational isomerization. Interestingly, the first and second phases appeared to be coupled since the amplitude of the second phase increased at the expense of the amplitude of the first phase in increasing urea concentrations. This observation together with the roll-over effect in the second folding phase indicates the presence of intermediate state during the folding reaction of HubWA. Quantitative analysis of Hub-WA folding kinetics indicated that this intermediate state is on the folding pathway. Folding kinetics measurement of a mutant HubWA with hydrophobic core residue mutation, Val to Ala at residue position 17, suggested that the intermediate state has significant amount of native interactions, supporting the interpretation that the intermediate is on the folding pathway. It is considered that HubWA is a useful model protein to study the contribution of residues to protein folding process using folding kinetics measurements in conjunction with protein engineering.

Expression level and glycan dynamics determine the net effects of TIMP-1 on cancer progression

  • Kim, Yong-Sam;Kim, Sun-Hee;Kang, Jeong-Gu;Ko, Jeong-Heon
    • BMB Reports
    • /
    • 제45권11호
    • /
    • pp.623-628
    • /
    • 2012
  • Tissue inhibitor of metalloproteinases (TIMPs; TIMP-1, -2, -3 and -4) are endogenous inhibitor for matrix metalloproteinases (MMPs) that are responsible for remodeling the extracellular matrix (ECM) and involved in migration, invasion and metastasis of tumor cells. Unlike under normal conditions, the imbalance between MMPs and TIMPs is associated with various diseased states. Among TIMPs, TIMP-1, a 184-residue protein, is the only N-linked glycoprotein with glycosylation sites at N30 and N78. The structural analysis of the catalytic domain of human stromelysin-1 (MMP-3) and human TIMP-1 suggests new possibilities of the role of TIMP-1 glycan moieties as a tuner for the proteolytic activities by MMPs. Because the TIMP-1 glycosylation participate in the interaction, aberrant glycosylation of TIMP-1 presumably affects the interaction, thereby leading to pathogenic dysfunction in cancer cells. TIMP-1 has not only the cell proliferation activities but also anti-oncogenic properties. Cancer cells appear to utilize these bilateral aspects of TIMP-1 for cancer progression; an elevated TIMP-1 level exerts to cancer development via MMP-independent pathway during the early phase of tumor formation, whereas it is the aberrant glycosylation of TIMP-1 that overcome the high anti-proteolytic burden. The aberrant glycosylation of TIMP-1 can thus be used as staging and/or prognostic biomarker in colon cancer.

In silico Analysis of Natural Compounds as Modulators of Type I Collagen

  • Narayanaswamy, Radhakrishnan;Wai, Lam Kok;Esa, Norhaizan Mohd;Ismail, Intan Safinar
    • 통합자연과학논문집
    • /
    • 제9권3호
    • /
    • pp.166-170
    • /
    • 2016
  • Collagen plays a vital role in the maintenance of structure and function of a human body. It has been widely applied in various fields including biomedical, cosmeceutical, food, pharmaceutical and tissue engineering. In the present study, the docking behaviour of type I collagen with 15 different ligands namely hydroxymethylfurfural, methylglyoxal, methylsyringate, O-methoxyacetophenone, 3-phenyllactic acid, 4-hydroxybenzoic acid, kojic acid, lumichrome, galangin, artoindonesianin F, caffeic acid, 4-coumaric acid, origanol A, thymoquinone and quercetin was evaluated along with their putative binding sites using Discovery Studio Version 3.1. Docking studies and binding free energy calculations revealed that origanol A has maximum interaction energy (-40.48 kcal/mol) and quercetin with the least interaction energy (-15.44 kcal/mol) as compared to the other investigated ligands. Three ligands which are galangin, methylsyringate and origanol A were shown to interact with Asp21 amino acid residue of chain B (type I collagen). Therefore, it is strongly suggested that the outcomes from the present study might provide new insight in understanding these 15 ligands as potential type I collagen modulators for the prevention of collagen associate disorders.

Digestate residues analysis under elevated heat regime by using DNS method

  • Hanif, Mohamad Haziq;Kamaruddin, Mohamad Anuar;Norashiddin, Faris Aiman;Zawawi, Mohd Hafiz
    • Advances in environmental research
    • /
    • 제9권1호
    • /
    • pp.55-64
    • /
    • 2020
  • The problems with unsorted municipal waste are always associated with disposal issues as it requires a large area for landfilling or high energy used for incineration. In recent years, an autoclaving technique has been considered a promising approach which could minimize the volume of organic waste from being directly disposed or incinerated. In this work, an attempt was done to study the saccharification potential of organic residues under elevated temperature Thermal treatment involving hot water bath was applied to treat the organic residue ranging from 60℃ to 100℃ for 30 and 60 minutes. The result obtained showed an increasing trend for the concentration of glucose and carbohydrate. However, the result for lignocellulose content which contains various component includes extractive, holocellulose, hemicellulose, cellulose and lignin show variation. Based on the thermal treatment carried out, the result indicated that the trend of glucose and carbohydrate content. The highest percentage of glucose that can be obtained 978.602 ㎍/ml which could be obtained at 90℃ at 60 minutes. The carbohydrate also shows an increasing trend with 0.234 mg/ml as the highest peak achieved at 80℃ for 30 minutes treatment. However, it was found that the lignocellulose content varies with temperature and time. The statistical analysis was carried out using two-ways ANOVA shows an interaction effect between the independent variables (temperature and contact time) and the saccharification effects on the food wastes. The result shows a variation in the significant effect of independent variables on the changes in the composition of food waste.

SPR-based Antibody-Antigen Interaction for Real Time Analysis of Carbamate Pesticide Residues

  • Yang, Gil-Mo;Kang, Suk-Won
    • Food Science and Biotechnology
    • /
    • 제17권1호
    • /
    • pp.15-19
    • /
    • 2008
  • This research was conducted to develop a quick and sensitive method of detecting carbamate residues using the immobilization of antibody-antigen interactions with surface plasmon resonance (SPR). We have used commercialized surface plasmon resonance equipment (Biacore 3000). The antibody used for the immunoassay was specific for glutathione-s-transferase (GST) and the antigens included several carbamate pesticides (carbofuran, carbaryl, and benfuracarb). When antigens were applied to the protein GST, the detection limit was 2 ng/mL of carbamate pesticide. The fabricated protein GST maintained its activity for over 200 measurements. Thus we determined that the SPR biosensors could detect the specific reversible binding of a reactant in solution to a binding partner immobilized on the surface of the sensor and allow real-time detection and monitoring.

Steady State Design for the Separation of Acetone-Chloroform Maximum Boiling Azeotrope Using Three Different Solvents

  • Pokhrel, Manish;Owusu, Asante Daniel;Cho, Jungho
    • Korean Chemical Engineering Research
    • /
    • 제55권4호
    • /
    • pp.490-496
    • /
    • 2017
  • We have designed an extractive distillation for separating maximum boiling azeotrope of acetone-chloroform system. PRO/II 9.4 was used to simulate the overall process. The VLE data adopted from Dortmund data bank was regressed to obtain a new set of binary interaction parameters. Three different entrainers were used for the separation process--dimethyl sulfoxide (DMSO), ethylene glycol (EG) and benzene--to test their viability for the acetone-chloroform system. Thermodynamic feasibility analysis was done through ternary map diagrams. Two different thermodynamic models, NRTL and UNIQUAC, were explored for the study of overall process.

Development, Validation, and Application of a Portable SPR Biosensor for the Direct Detection of Insecticide Residues

  • Yang, Gil-Mo;Cho, Nam-Hong
    • Food Science and Biotechnology
    • /
    • 제17권5호
    • /
    • pp.1038-1046
    • /
    • 2008
  • This study was carried out to develop a small-sized biosensor based on surface plasmon resonance (SPR) for the rapid identification of insecticide residues for food safety. The SPR biosensor module consists of a single 770 nm-light emitting diodes (LED) light source, several optical lenses for transferring light, a hemisphere sensor chip, photo detector, A/D converter, power source, and software for signal processing using a computer. Except for the computer, the size and weight of the sensor module are 150 (L)$\times$70 (W)$\times$120 (H) mm and 828 g, respectively. Validation and application procedures were designed to assess refractive index analysis, affinity properties, sensitivity, linearity, limits of detection, and robustness which includes an analysis of baseline stability and reproducibility of ligand immobilization using carbamate (carbofuran and carbaryl) and organophosphate (cadusafos, ethoprofos, and chlorpyrifos) insecticide residues. With direct binding analysis, insecticide residues were detected at less than the minimum 0.01 ppm and analyzed in less than 100 sec with a good linear relationship. Based on these results, we find that the binding interaction with active target groups in enzymes using the miniaturized SPR biosensor could detect low concentrations which satisfy the maximum residue limits for pesticide tolerance in Korea, Japan, and the USA.

Mutagenic Analysis of hPNMT Confirms the Importance of Lys57 and the Inhibitor Binding Site

  • Jeong, Ki-Woong;Kang, Dong-Il;Lee, Jee-Young;Kim, Yang-Mee
    • Bulletin of the Korean Chemical Society
    • /
    • 제32권2호
    • /
    • pp.455-458
    • /
    • 2011
  • In previous report, with the aid of receptor-oriented pharmacophore-based in silico screening, we characterized three novel hPNMT inhibitors (YPN010, YPN016, and YPN017) and proposed that the hydrogen bonding interaction between inhibitors and side chain of Lys57 is very important to inhibitory activity of hPNMT. To confirm the importance of Lys57, mutant with substitution of Lys57 with Ala was cloned and binding study was performed for a K57A mutant of hPNMT using STD-NMR and fluorescence experiments. The binding constants for three novel inhibitors with mutant hPNMT were dramatically decreased compared to those with wild-type protein. K57A mutant-induced conversion of noradrenaline to adrenaline was suppressed about 95 % compared to wild-type hPNMT. Mutagenic analysis using a K57A mutant confirmed the importance of the Lys57 residue in binding of the inhibitor candidate to hPNMT as well as enzymatic activity of hPNMT, implying that these results are consistent with our binding model.