• Title/Summary/Keyword: protein displacement

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Dynamics of a Globular Protein and Its Hydration Water Studied by Neutron Scattering and MD Simulations

  • Kim, Chan-Soo;Chu, Xiang-Qiang;Lagi, Marco;Chen, Sow-Hsin;Lee, Kwang-Ryeol
    • Proceedings of the Korean Vacuum Society Conference
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    • 2011.02a
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    • pp.21-21
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    • 2011
  • A series of Quasi-Elastic Neutron Scattering (QENS) experiments helps us to understand the single-particle (hydrogen atom) dynamics of a globular protein and its hydration water and strong coupling between them. We also performed Molecular Dynamics (MD) simulations on a realistic model of the hydrated hen-egg Lysozyme powder having two proteins in the periodic box. We found the existence of a Fragile-to-Strong dynamic Crossover (FSC) phenomenon in hydration water around a protein occurring at TL=$225{\pm}5K$ by analyzing Intermediate Scattering Function (ISF). On lowering of the temperature toward FSC, the structure of hydration water makes a transition from predominantly the High Density Liquid (HDL) form, a more fluid state, to predominantly the Low Density Liquid (LDL) form, a less fluid state, derived from the existence of a liquid?liquid critical point at an elevated pressure. We showed experimentally and confirmed theoretically that this sudden switch in the mobility of the hydration water around a protein triggers the dynamic transition (so-called glass transition) of the protein, at a temperature TD=220 K. Mean Square Displacement (MSD) is the important factor to show that the FSC is the key to the strong coupling between a protein and its hydration water by suggesting TL${\fallingdotseq}$TD. MD simulations with TIP4P force field for water were performed to understand hydration level dependency of the FSC temperature. We added water molecules to increase hydration level of the protein hydration water, from 0.30, 0.45, 0.60 and 1.00 (1.00 is the bulk water). These confirm the existence of the FSC and the hydration level dependence of the FSC temperature: FSC temperature is decreased upon increasing hydration level. We compared the hydration water around Lysozyme, B-DNA and RNA. Similarity among those suggests that the FSC and this coupling be universal for globular proteins, biopolymers.

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Evidence for Regulation of Interaction of Endogenous Protein Kinase C(Pkc) Substrates with Plasma Membrane by PKC Down-Regulation in K562 Cells

  • Kim, Young-Sook
    • Archives of Pharmacal Research
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    • v.18 no.5
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    • pp.301-307
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    • 1995
  • In the particulate fraction obtained from PKC-down regulated K562 cells by treatment for 24 h with 200nM TPA, phosphorylation of two proteins with molecular weight, 100 kDa and 23 kDa (designated p100 and p23, respectvely) was depleted and addition of exogenous purified PKC to this fraction failed to testore their phosphorylation. However, in the soluble fraction, all of phosphoproteins abolished by long-term treatment with TPA were restored by exogenously added PKC. Phosphorylation of two proteins was increased by short-term tretment (20 min), and diminished with the persistant exposure to TPA as well as at a concentration as low as 1nM. When K562 cells were treated with 1 nM and 200 nM TPA for 24 h, phosphorylation of p100 was restored with or without exogenous PKC on 2-3day and 6day after removal of treated TPA, respectively. Two-dimensional electrophoresis of phosphoproteins revealed that phosphorylated p100 (pl=5.9) and p66 species were completely absent from the particulate fraction of K562 cells treated with 200nM TPA for 24 h. These results suggest that the interaction of sensitive endogenous substrates, p100 and p23 with the plasma membrane might be regulated by PKC-down regulation without displacement to the cytosol and the interaction of p100 with the membrane might be reveersible.

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Surface Composition and Molecular Diffusion on the Stability of Foams Formed from Protein/Surfactant Mixtures (단백질/계면활성제 혼합계에서 기포안정성에 대한 계면조성과 분자확산)

  • Park, Sun-Yeong;Kim, Myung-Soo;Jeong, Noh-Hee;Nam, Ki-Dae
    • Journal of the Korean Applied Science and Technology
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    • v.17 no.3
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    • pp.158-166
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    • 2000
  • A conductimetric study of foam formed from mixture of the protein, ${\beta}-lactoglobulin$, and the nonioinc surfactant, SML, revealed that their stability was reduced at concentrations of SML in the range $3{\sim}10mM$. The interaction of SML with ${\beta}-lactoglobulin$ was investigated by fluorimetry and a dissociation constant of $0.2{\mu}M$ was calculated for the complex. Surface tension studies confirmed the presence of interaction between the two components and provided evidence for the progressive displacement of ${\beta}-lactogloblin$ from the air/water interface with increasing SML concentration. Experiments using air-suspended microscopic thin liquid films revealed transitions in the chainage characteristics and thickness of the film at SML concentrations below that which resulted in destabilization of the foam. However, measurements of surface mobility of fluorescent-labeled ${\beta}-lactoglobulin$ by a photobleaching method identified that a transition to a mobile system occurred at a SML concentration which correlated with the onset of instability in the disperse phase. The results would indicate that maintenance of the viscoelastic properties of the surface is paramount importance in determining the stability of interfaces comprising mixtures of protein and surfactant.

The nano scale bending and dynamic properties of isolated protein microtubules based on modified strain gradient theory

  • Benmansour, Djazia Leila;Kaci, Abdelhakim;Bousahla, Abdelmoumen Anis;Heireche, Houari;Tounsi, Abdelouahed;Alwabli, Afaf S.;Alhebshi, Alawiah M.;Al-ghmady, Khalid;Mahmoud, S.R.
    • Advances in nano research
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    • v.7 no.6
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    • pp.443-457
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    • 2019
  • In this investigation, dynamic and bending behaviors of isolated protein microtubules are analyzed. Microtubules (MTs) can be considered as bio-composite structures that are elements of the cytoskeleton in eukaryotic cells and posses considerable roles in cellular activities. They have higher mechanical characteristics such as superior flexibility and stiffness. In the modeling purpose of microtubules according to a hollow beam element, a novel single variable sinusoidal beam model is proposed with the conjunction of modified strain gradient theory. The advantage of this model is found in its new displacement field involving only one unknown as the Euler-Bernoulli beam theory, which is even less than the Timoshenko beam theory. The equations of motion are constructed by considering Hamilton's principle. The obtained results are validated by comparing them with those given based on higher shear deformation beam theory containing a higher number of variables. A parametric investigation is established to examine the impacts of shear deformation, length scale coefficient, aspect ratio and shear modulus ratio on dynamic and bending behaviors of microtubules. It is remarked that when length scale coefficients are almost identical of the outer diameter of MTs, microstructure-dependent behavior becomes more important.

Correlation between Rab3A Expression and Sperm Kinematic Characteristics

  • Seung-Ik Jang;Jae-Hwan Jo;Uwamahoro Claudine;Eun-Ju Jung;Woo-Jin Lee;Ju-Mi Hwang;Jeong-Won Bae;Dae-Hyun Kim;Jun Koo Yi;Jae Jung Ha;Dong Yep Oh;Woo-Sung Kwon
    • Development and Reproduction
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    • v.28 no.1
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    • pp.13-19
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    • 2024
  • Ras-related (Rab) proteins, integral members of the monomeric G-protein family, play a pivotal role in regulating intracellular vesicular transport. These proteins contribute to male reproductive processes, specifically in acrosome formation, exocytosis, and sperm motility. Although a prior study indicated a correlation between Rab3A and sperm motility, including motion kinematic parameters such as mean dance, this association has only been explored within a limited sample size. Therefore, further verification is required to confirm the correlation between Rab3A and sperm motility parameters. In the present study, Rab3A expression, sperm motility, and motion kinematic parameters were analyzed in 150 boar spermatozoa. Additionally, correlations between Rab3A expression and sperm kinematic characteristics were evaluated statistically. The results revealed significant associations between Rab3A protein expression levels and various motion kinematic parameters. Specifically, Rab3A levels exhibited positive correlations with average path velocity (p<0.05), mean amplitude of lateral head displacement (p<0.05), and curvilinear velocity (p<0.01). Consequently, it is proposed that Rab3A protein plays a crucial role in male fertility through its correlation with sperm kinematic characteristics, making it a potential marker for sperm motility-related assessments.

Aptamer-based optical switch for biosensors (압타머 광학 바이오센서)

  • Lee, Joo-Woon;Cho, Jeong Hwan;Cho, Eun Jeong
    • Analytical Science and Technology
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    • v.27 no.3
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    • pp.121-139
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    • 2014
  • In this review, we will discuss aptamer technologies including in vitro selection, signal transduction mechanisms, and designing aptamers and aptazyme for label-free biosensors and catalysts. Dye-displacement, a typical label-less method, is described here which allows avoiding relatively complex labeling steps and extending this application to any aptamers without specific conformational changes, in a more simple, sensitive and cost effective way. We will also describe most recent and advanced technologies of signaling aptamer and aptazyme for the various analytical and clinical applications. Quantum dot biosensor (QDB) is explained in detail covering designing and adaptations for multiplexed protein detection. Application to aptamer array utilizing self-assembled signaling aptamer DNA tile and the novel methods that can directly select smart aptamer or aptazyme experimentally and computationally will also be finally discussed, respectively.

Analysis of Low Velocity Impact on Biomimetic Composites Mimicking Nacre (진주조개를 모방한 생체모방 복합재료의 저속충격 해석)

  • Jo, Seung-Un;Beom, Hyeon-Gyu
    • Composites Research
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    • v.23 no.4
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    • pp.1-6
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    • 2010
  • The dynamicresponse of biomimetic composites mimicking nacre under low velocity impact is investigated. The composites have hierarchical structures with a staggered pattern consisting of a protein and a mineral. To analyze the impact response of the composites, the finite element method is used. The effects of the hierarchical structures of the compositeson the dynamic response are examined. It is shown that the maximum stress, displacement and contact force in the composite subjected to low velocity impact decrease as the level of structural hierarchy increases.

Sequencing and annotation of the complete mitochondrial genome of a threatened labeonine fish, Cirrhinus reba

  • Islam, Mohammad Nazrul;Sultana, Shirin;Alam, Md. Jobaidul
    • Genomics & Informatics
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    • v.18 no.3
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    • pp.32.1-32.7
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    • 2020
  • The mitochondrial genome of a species is an essential resource for its effective conservation and phylogenetic studies. In this article, we present sequencing and characterization of the complete mitochondrial genome of a threatened labeonine fish, Cirrhinus reba collected from Khulna region of Bangladesh. The complete mitochondrial genome was 16,597 bp in size, which formed a circular double-stranded DNA molecule containing a total of 37 mitochondrial genes (13 protein-coding genes, 2 ribosomal RNA genes, and 22 transfer RNA genes) with two non-coding regions, an origin of light strand replication (OL) and a displacement loop (D-loop), similar structure with other fishes of Teleostei. The phylogenetic tree demonstrated its close relationship with labeonine fishes. The complete mitogenome of Cirrhinus reba (GenBank no. MN862482) showed 99.96% identity to another haplotype of Cirrhinus reba (AP013325), followed by 90.18% identity with Labeo bata (AP011198).

Whole Food Interventions and Nutrient Displacement: Tomato Sauce Entrees for Prostate Cancer Patients

  • Hwang, Eun-Sun;Duncan, Claudine E.;Bowen, Phyllis E.
    • Preventive Nutrition and Food Science
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    • v.14 no.4
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    • pp.343-348
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    • 2009
  • This study sought to determine changes in self-selected diets in response to a high energy tomato pasta entree. Thirty men, mostly African-American, who were diagnosed with prostate cancer and also scheduled for prostatectomy were enrolled in the study. Dietary intakes were obtained by 24 hr diet recall for 3 days before the intervention and 3 days in week 2 during the intervention. Tomato pasta entrees were formulated to contain 30 mg of lycopene with roughly the same macronutrient composition and averaged 771 kcal/entree. Mean adherence to lycopene dose was 82% and, days when the entree was consumed, the mean adherence to lycopen was 90%. Lycopene intake in their self-selected diet decreased from 5 to 1 mg/day which lowered the exposure to lycopene, planned during the intervention. The men were able to decompensate for most of the added energy to their diet, but there was still a mean increase in total intake of 242 kcal/day (p=0.04), which did not result in weight gain over the short period of the study. There were no significant changes in the percent of energy from protein, fat or carbohydrates, but dietary cholesterol increased from 341 to 472 mg/day (p=0.0002). Mean intakes of vitamins A and C, and folate were not significantly different and were above the EAR. Vitamin E intake decreased, but, because of the variance in intake, the decrease was not statistically significant. Possible deterioration of diet quality should be a consideration when recommending whole food interventions for the prevention of chronic disease or the amelioration of physiological dysfunction.

Development of a Sandwich Enzyme Immunoassay for Salmon Gonadotropin II. (연어 생식소자극호르몬 II의 Sandwich Enzyme Immunoassay법 개발)

  • KIM Dae-Jung;HAN Chang-Hee;AIDA Katsumi
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.33 no.1
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    • pp.55-59
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    • 2000
  • A specific and sensitive sandwich enzyme-immunoassay (EIA) using Avidin-Biotin complex was developed for the measurement of GTH II levels in pituitary content and pituitary cell culture medium of the rainbow trout-(Oncorhpchus mykiss). Biotin-salmon GTH II rabbit IgG (sefondary antibody) wai purified by a protein A sepharose affinity chromatography column and that was biotinylated by using Biotin-N-hydroxysuccinimide ofter (BNHS). Non-biotin salmon GTH II rabbit IgG (first antibody) was obtained only through a protein A sepharose affinity chromatography column. The assay was performed by the so-called 'sandwich' method using a microtiter plate, A dose-response curve was obtained between $0.12 to 125 ng/ml$ of salmon GTH II. The displacement curves for pituitary extraction and pituitary cell culture medium of testosterone-treated rainbow trout were Parallel to the standard curie. The intra-assay and inter-assay coefficients of variation (CV) were $8.2{\%} (N=5) and 12.5{\%} (N=6)$, respectively, This assay system was used to measure the amount of GTH II that accumulated in the culture medium of dispersed pituitary cells in testosterone-treated immature rainbow trout, The accumulation was increased with the amount or salmon gonadotropin-releasing hormone. GTH II values determined by the present method were well correlated with those determined by radioimmunoassay. As a result, this assay system was found to be suitable for the measurement of GTH II for pituitary extraction and pituitary culture medium in many salmonid fishes.

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