• Title/Summary/Keyword: Protein sensor

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Determination of $K^+-, Ca^{2}+- and Mg^{2+}-$ATPase activities in Fish Muscle Protein by ATPase Biosensor (ATPase 센서를 이용한 어류근육 단백질의 $K^+-, Ca^{2}+- 및 Mg^{2+}-$ ATPase 활성의 측정)

  • 천병수;김희경
    • KSBB Journal
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    • v.11 no.5
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    • pp.518-523
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    • 1996
  • The sensor to determine ATPase activities was consisted of an immobilized enzyme membrane(purine nucloside phosphoryrase and xanthine oxidase) and an oxygen electrode. The proposed sensor was used for the determination of $K^+-, Ca^{2}+- and Mg^{2+}-$ATPase activities in several fish muscle proteins such as Thunnus albacares(Yellowfin tuna), Tetrapturus audax(Striped marlin), Prognichthys agoo(Japanese flyingfish), and Cypvinus carpio(Carp). $K^+-, Ca^{2}-$ATPase activities measured by the proposed sensor system were in good agreement with the results obtained by a conventional colorimetric assay. One cycle of assay could be completed within 3mlnutes.

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Establishment of a NanoBiT-Based Cytosolic Ca2+ Sensor by Optimizing Calmodulin-Binding Motif and Protein Expression Levels

  • Nguyen, Lan Phuong;Nguyen, Huong Thi;Yong, Hyo Jeong;Reyes-Alcaraz, Arfaxad;Lee, Yoo-Na;Park, Hee-Kyung;Na, Yun Hee;Lee, Cheol Soon;Ham, Byung-Joo;Seong, Jae Young;Hwang, Jong-Ik
    • Molecules and Cells
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    • v.43 no.11
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    • pp.909-920
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    • 2020
  • Cytosolic Ca2+ levels ([Ca2+]c) change dynamically in response to inducers, repressors, and physiological conditions, and aberrant [Ca2+]c concentration regulation is associated with cancer, heart failure, and diabetes. Therefore, [Ca2+]c is considered as a good indicator of physiological and pathological cellular responses, and is a crucial biomarker for drug discovery. A genetically encoded calcium indicator (GECI) was recently developed to measure [Ca2+]c in single cells and animal models. GECI have some advantages over chemically synthesized indicators, although they also have some drawbacks such as poor signal-to-noise ratio (SNR), low positive signal, delayed response, artifactual responses due to protein overexpression, and expensive detection equipment. Here, we developed an indicator based on interactions between Ca2+-loaded calmodulin and target proteins, and generated an innovative GECI sensor using split nano-luciferase (Nluc) fragments to detect changes in [Ca2+]c. Stimulation-dependent luciferase activities were optimized by combining large and small subunits of Nluc binary technology (NanoBiT, LgBiT:SmBiT) fusion proteins and regulating the receptor expression levels. We constructed the binary [Ca2+]c sensors using a multicistronic expression system in a single vector linked via the internal ribosome entry site (IRES), and examined the detection efficiencies. Promoter optimization studies indicated that promoter-dependent protein expression levels were crucial to optimize SNR and sensitivity. This novel [Ca2+]c assay has high SNR and sensitivity, is easy to use, suitable for high-throughput assays, and may be useful to detect [Ca2+]c in single cells and animal models.

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

  • Yang, Gil-Mo;Kang, Suk-Won
    • Food Science and Biotechnology
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    • v.17 no.1
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    • pp.15-19
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    • 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.

AMP-activated protein kinase: An emerging target for ginseng

  • Jeong, Kyong Ju;Kim, Go Woon;Chung, Sung Hyun
    • Journal of Ginseng Research
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    • v.38 no.2
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    • pp.83-88
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    • 2014
  • The adenosine monophosphate (AMP)-activated protein kinase (AMPK) is a key sensor of cellular energy. Once activated, it switches on catabolic pathways generating adenosine triphosphate (ATP), while switching off biosynthetic pathways consuming ATP. Pharmacological activation of AMPK by metformin holds a therapeutic potential to reverse metabolic abnormalities such as type 2 diabetes and nonalcoholic fatty liver disease. In addition, altered metabolism of tumor cells is widely recognized and AMPK is a potential target for cancer prevention and/or treatment. Panax ginseng is known to be useful for treatment and/or prevention of cancer and metabolic diseases including diabetes, hyperlipidemia, and obesity. In this review, we discuss the ginseng extracts and ginsenosides that activate AMPK, we clarify the various mechanisms by which they achieve this, and we discuss the evidence that shows that ginseng or ginsenosides might be useful in the treatment and/or prevention of metabolic diseases and cancer.

Expression of Beta-catenin-interacting Protein 1 (CTNNBIP1) Gene Is Increased under Hypothermia but Decreased under Additional Ischemia Conditions

  • Kwon, Kisang;Kim, Seung-Whan;Yu, Kweon;Kwon, O-Yu
    • Biomedical Science Letters
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    • v.20 no.3
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    • pp.168-172
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    • 2014
  • It has recently been shown that hypothermia treatment improves brain ischemia injury and is being increasingly considered by many clinicians. However, the precise roles of hypothermia for brain ischemia are not yet clear. In the present study we demonstrated firstly that hypothermia induced beta-catenin-interacting protein 1 (CTNNBIP1) gene expression and its expression was dramatically decreased under ischemic conditions. It was also demonstrated that hypothermia activated endoplasmic reticulum (ER) stress sensors especially both, the phosphorylation of $eIF2{\alpha}$, and ATF6 proteolytic cleavage. However, the factors of apoptosis and autophagy were not associated with hypothermia. These findings suggested that hypothermia controlled CTNNBIP1 gene expression under ischemia, which may provide a clue to the development of treatments and diagnostic methods for brain ischemia.

c-myc Expression: Keep the Noise Down!

  • Chung, Hye-Jung;Levens, David
    • Molecules and Cells
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    • v.20 no.2
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    • pp.157-166
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    • 2005
  • The c-myc proto-oncogene encodes a nuclear protein that is deregulated and/or mutated in most human cancers. Acting primarily as an activator and sometimes as a repressor, MYC protein controls the synthesis of up to 10-15% of genes. The key MYC targets contributing to oncogenesis are incompletely enumerated and it is not known whether pathology arises from the expression of physiologic targets at abnormal levels or from the pathologic response of new target genes that are not normally regulated by MYC. Regardless of which, available evidence indicates that the level of MYC expression is an important determinant of MYC biology. The c-myc promoter has architectural and functional features that contribute to uniform expression and help to prevent or mitigate conditions that might otherwise create noisy expression. Those features include the use of an expanded proximal promoter, the averaging of input from dozens of transcription factors, and real-time feedback using the supercoil-deformable Far UpStream Element (FUSE) as physical sensor of ongoing transcriptional activity, and the FUSE binding protein (FBP) as well as the FBP interacting repressor (FIR) as effectors to enforce normal transcription from the c-myc promoter.

재조합 Escherichia coli에 의한 5-Aminolevulinic acid (ALA) 생산 연구

  • Seo, Guk-Hwa;Lee, Jong-Il
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.715-718
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    • 2001
  • 5-Aminolevulinic acid (ALA) can be used as a photodynamic herbicide / insecticide and also applied to photodynamic therapy for malignant skin tumor. In this study we developed recombinant E. coli system for the high productivity of ALA and for the on-line monitoring of fusion protein (EGFP::ALAS) by a fluorescence sensor.

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Endoplasmic Reticulum Signaling for Recombinant-protein Production (재조합 단백질 생산을 위한 소포체 신호전달)

  • Goo, Tae-Won;Yun, Eun-Young;Kang, Seok-Woo;Kwon, Ki-Sang;Kwon, O-Yu
    • Journal of Life Science
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    • v.17 no.6 s.86
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    • pp.847-858
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    • 2007
  • The endoplasmic reticulum (ER) is an important intracellular organelle for folding and maturation of newly synthesized transmembrane and secretory proteins. The ER provides stringent quality control systems to ensure that only correctly folded proteins exit the ER and unfolded or misfolded proteins are retained and ultimately degraded. The ER has evolved stress response both signaling pathways the unfolded protein response (UPR) to cope with the accmulation of unfolded or misfolded proteins and ER overload response (EOR). Accumulating evidence suggests that, in addition to responsibility for protein processing, ER is also an important signaling compartment and a sensor of cellular stress. In this respect, production of bio-functional recombinant-proteins requires efficient functioning of the ER secretory pathway in host cells. This review briefly summarizes our understanding of the ER signaling developed in the recent years to help of the secretion capacities of recombinant cells.

ANALYSIS OF FLUIDIC BEAD CUBE EMBEDDED PORTABLE CMOS SENSING SYSTEM FOR IMMUNO REACTION MONITORING (유체소자가 집적화된 면역검사용 휴대용 CMOS 바이오칩의 분석)

  • Jeong, Yong-Won;Park, Se-Wan;Kim, Jin-Seok;Kim, Hyeon-Cheol;Chun, Kuk-Jin
    • Proceedings of the IEEK Conference
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    • 2005.11a
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    • pp.755-758
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    • 2005
  • This paper describes the novel immunoassay sensing system for a portable clinical diagnosis system. It consists of a bead cage reactor and a CMOS integrated biosensor. It showed the simple and easy antibody coating method on beads by flow-through avidin biotin complex technology in a microfluidic device. It showed just 90 nL sample consumption and good result for the application of alpha feto protein. The bead cage reactor has the role of the antibody coating, antigen binding and enzyme linking for the electrochemical sensing method. The CMOS biosensor consists of ISFET (ion selective field effect transistor) biosensor and temperature sensor for detecting pH that is the byproduct of enzyme reaction. The sensitivity is 8 $kHz/^{\circ}C$ in a temperature sensor and 33 mV/pH in a pH sensor. After filling the 15 um polystyrene beads in bead cage, antibody flowed and reacted to beads. Subsequently, the biotinylated antigen flowed and bound to the antibody and GOD (glucose oxidase)-avidin conjugate flowed and reacted to the biotin of the biotinylated antigen. After this reaction process, glucose solution flowed and reacted to the GOD on beads. The hydrogen was generated by glucose-GOD reaction. And it was detected by the pH sensor.

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BIOSENSORS IN AGRICULTURAL AND BIOLOGICAL SYSTEMS (농업 및 생물계에서의 바이오 센서)

  • Delwiche, M.J.;Jenkins, D.M.;Tang, X.;Jackson, E.S.
    • Proceedings of the Korean Society for Agricultural Machinery Conference
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    • 2000.11a
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    • pp.76-100
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    • 2000
  • A brief general discussion of the nature and function of biosensors is presented. While the primary motivator for biosensor development has been the health-care industries, recent research efforts have spread to problems in agriculture and biological production systems. To illustrate some of the research from our laboratory, three example biosensors and their corresponding applications are presented. The first of these is an immunosensor for measurement of the hormone progesterone during milking as a method to improve reproductive management of dairy herds. The second example is an enzyme sensor for measurement of urea in milk as a menas to determine the efficiency of conversion of input protein to milk protein and, thus, improve nutritional management of dairy herds. The third example is a DNA sensor using polymerase chain reaction to detect pathogenic bacteria in the wash water of fresh and minimally processed fruits and vegetables. The potential for application of biosensors in agriculture, agrobiotechnology, food processing, and environmental monitoring has barely been realized.

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