• 한국환경성돌연변이발암원학회:학술대회논문집
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• 한국환경성돌연변이발암원학회 2002년도 Current Trends in Toxicological Sciences
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• pp.25-35
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• 2002

Transcription factor Nrf2 is essential for the antioxidant responsive element (ARE)-mediated induction of phase II detoxifying and oxidative stress enzyme genes. Detailed analysis of differential Nrf2 activity displayed in transfected cell lines ultimately led to the identification of a new protein, which we named Keap1, that suppresses Nrf2 transcriptional activity by specific binding to its evolutionarily conserved amino-terminal regulatory domain. The closest homolog of Keap1 is a Drosophila actin-binding protein called Kelch, implying that Keap1 might be a Nrf2 cytoplasmic effector. We then showed that electrophilic agents antagonize Keap1 inhibition of Nrf2 activity in vivo, allowing Nrf2 to traverse from the cytoplasm to the nucleus and potentiate the ARE response. We postulate that Keap1 and Nrf2 constitute a crucial cellular sensor for oxidative stress, and together mediate a key step in the signaling pathway that leads to transcriptional activation by this novel Nrf2 nuclear shuttling mechanism. The activation of Nrf2 leads in turn to the induction of phase II enzyme and antioxidative stress genes in response to electrophiles and reactive oxygen species.

• Applied Biological Chemistry
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• 제49권3호
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• pp.254-258
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• 2006

내분비계장애물질에 노출된 어류의 에스트로겐 활성에 대한 생물지표단백질인 vitellogenin을 검출하는 압전류적 면역센서를 잉어 vitellogenin에 대한 항체와 AT-cut 수정진동자를 생물요소와 변환기로 사용하여 구성하고 이를 이용한 잉어 vitellogenin 검출을 행하였다. 이형이기능성의 티올화 가교화제인 sulfosuccinimidyl 6-[3-(2-pyridyldithio)propionamido]hexanoate 처리에 의하여 티올화된 항체를 수정진동자상의 금전극에 화학흡착법에 의하여 고정화하여 센서 chip을 제조하였다. 센서반응을 위한 반응완충용액을 0.1M sodium phosphate(pH 7.4)로 선정한 후 $0.4864{\sim}486.4000\;nM$의 vitellogenin 용액을 가하였을 때 농도의존적인 센서반응의 증가가 나타났으며 이 때 잉어 vitellogenin에 대한 검출한계는 0.4864 nM로 추정되었다.

• 한국콘텐츠학회논문지
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• 제21권4호
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• pp.825-832
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• 2021

본 연구는 치주질환의 주 원인균인 P. gingivalis에 선택적으로 작용하는 특이적 앱타머를 선별하고 선별된 앱타머와 결합하는 단백질 분자를 정제 및 동정함으로써 P. gingivalis에 관한 작용기전을 규명하고자 하였다. 이를 위하여 39개의 random sequence를 갖는 DNA library를 제조하여 SELEX 방법을 이용하여 P. gingivalis에 특이성을 가진 앱타머를 선별하였으며 PCR2.1 cloning vector를 활용한 cloning을 시행하여 염기서열을 분석했다. 8종의 각기 다른 염기서열을 가진 앱타머를 선별하였고 직접적으로 작용하는 단백질을 밝혀내고자 선별된 앱타머 중 APG-3를 이용하여 modified weston blot을 시행하고 단백질을 분석한 결과 P. gingivalis에 선택적으로 결합하는 11종의 단백질을 분리, 동정하였다. 이와 같은 결과로 앱타머가 치주질환의 원인균인 P. gingivalis의 당 대사 및 세포활성억제와 관련된 단백질에 선택적으로 결합하여 부착함으로써 치주질환의 진단을 위한 센서로 가능성을 제시했다.

• 한국미생물·생명공학회지
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• 제48권1호
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• pp.24-31
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• 2020

Five genes (mxbDM, mxcQE and mxaB) are responsible for the transcription of methanol oxidation genes in Methylobacterium strains. Among these, MxbDM and MxcQE constitute the two-component system (TCS) regulating methanol metabolism. In this study, we integrated the methanol-sensing domain of MxbD and MxcQ with the EnvZ/OmpR from Escherichia coli. The domain-swapping strategy resulted in chimeric histidine kinases (HK's) MxbDZ and MxcQZ AM1 containing recombinant E. coli. Real-time quantitative PCR was used to monitor OmpC expression mediated by the chimeric HK and response regulator (RR) OmpR. Further, an ompC promoter based fluorescent biosensor for sensing methanol was developed. GFP fluorescence was studied both qualitatively and quantitatively in response to environmental methanol. GFP measurement also confirmed ompC expression. Maximum fluorescence was observed at 0.05% methanol and 0.01% methanol using MxbDZ and MxcQZ AM1, respectively. Thus the chimeric HK containing E. coli were found to be highly sensitive to methanol, resulting in a rapid response making them an ideal sensor.

• BMB Reports
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• 제32권3호
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• pp.215-225
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• 1999

Phytochromes (with gene family members phyA, B, C, D, and E) are a wavelength-dependent light sensor or switch for gene regulation that underscore a number of photo responsive developmental and morphogenic processes in plants. Recently, phytochrome-like pigment proteins have also been discovered in prokaryotes, possibly functioning as an auto-phosphorylating/phosphate-relaying two-component signaling system (Yeh et al., 1997). Phytochromes are photochromically convertible between the light sensing Pr and regulatory active Pfr forms. Red light converts Pr to Pfr, the latter having a "switch-on" conformation. The Pfr form triggers signal transduction pathways to the downstream responses including the expression of photosynthetic and other growth-regulating genes. The components involved in and the molecular mechanisms of the light signal transduction pathways are largely unknown, although G-proteins, protein kinases, and secondary messengers such as $Ca^{2+}$ ions and cGMP are implicated. The 124-127 kDa phytochromes form homodimeric structures. The N-terminal half contains the tetrapyrrolic phytochromobilin for red/far-red light absorption. The C-terminal half includes both a dimerization motif and regulatory box where the red light signal perceived by the chromophore-domain is recognized and transduced to initiate the signal transduction cascade. A working model for the inter-domain signal communication within the phytochrome molecule is proposed in this Review.

• BMB Reports
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• 제30권5호
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• pp.308-314
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• 1997

We investigated biotinylation of nitrocellulose membrane for immuno-filtration capture assay. In order to enhance the efficiency of biotinylation, nitrocellulose membranes were pretreated with several chemicals for the purpose of suitable protein absorption through surface modification. As a signal generating enzyme, urease was used and the concentration of avidin was optimized for the efficient binding kinetics between urease-biotin in liquid phase and biotinylated membrane in solid phase. For effective biotinylation, bovine serum albumin-biotin complexes could be immobilized at a concentration of $370\;{\mu}g$/stick ($4.4\;cm^2$). Among tested chemicals, polylysine (0.25%) showed a significant effect in biotinylation. Polylysine is thought to enhance surface area by extending unbound residues into solution. Time of treatment over 30 min and higher molecular weight of polylysines (58,100 dalton) showed positive effect on the enhancement of biotinylation. The result from this study may be useful for developing a new biosensor and other biofunctional membranes for examining molecular recognition.

• 대한전기학회:학술대회논문집
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• 대한전기학회 2009년도 제40회 하계학술대회
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• pp.1545_1546
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• 2009

This paper reports a novel wireless love-wave biosensor on $41^{\circ}$ YX $LiNbO_3$ piezoelectric substrate and $SiO_2$ guiding layer for Immunoglobulin G (IgG) detection by protein binding. Different from the traditional biosensors based on surface acoustic wave (SAW) oscillator structured by delay line/resonators, a 440MHz reflective delay line consists of SPUDTs and three reflectors placed on $41^{\circ}$ YX $LiNbO_3$ in a row was fabricated as the sensor element. Good linearity, reproducibility, and high sensitivity were observed in the IgG concentration range 1~65nM. Unique advantages as high sensitivity, passive and simple measurement system are present over currently available other biosensors.

• 한국농업기계학회:학술대회논문집
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• 한국농업기계학회 1993년도 Proceedings of International Conference for Agricultural Machinery and Process Engineering
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• pp.1354-1363
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• 1993

Optical methods were developed to examine their feasibility for quality evaluation of peanut with pod. Surface color and internal quality of peanut were measured without contact. The surface color of peanut was measured by light reflectance at a region of visible wavelengths. Its characteristic was high correlated with a visual grading of peanut. A trial machine for the color grading of peanut was developed using an optical sensor and it was considered to compare with the visual grading. The spectral reflectance at a region of near infrared wavelengths from 1,200 to 2,500nm was measured , and the chemical components of peanut were related to spectral reflectance at special wavelengths. The protein, fat and moisture contents of peanut were estimated by the near infrared methods. An infrared imaging method was developed to evaluate the internal quality of peanut with pod. As thermal characteristic of peanut with pod was deeply related to internal quality , the quality of peanut can be evaluated by temperature changes on the surface of peanut. Measurement of surface color, near infrared reflectance and thermal imaging were shown to be very effective in grading of peanut with pod.

• 한국가시화정보학회:학술대회논문집
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• 한국가시화정보학회 2002년도 마이크로/바이오 가시화기술부문 학술강연회
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• pp.1-7
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• 2002

This papers describes the preparation and experimental results of a micro mass detection devices based on cantilever and a diffuser-type micro pump using screen printing thick-film technologies and Si micro-machining. PZT-PCW thick films were prepared by new hybrid method based on the screen printing. By applying these PZT-PCW piezoelectric thick films on actuator, a cantilever for mass detection sensor and a micropump for microfluidic element are successfully fabricated. Resonant frequency and displacement of PZT-PCW thick film actuator in air and in liquid are measured by laser vibrometer system as a function of actuator size. The resonant frequency of PZT-PCW thick film actuator in liquid decreases order of 1/2-1/4 due to damping effect. The sensitivity of cantilever is characterized by Au deposition method which has the mass loading effect such as adsorption of protein. The Sensitivity of PZT-0.12PCW thick film cantilever is proportional to detecting area.

• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2005년도 추계학술대회 및 한일 식물생명공학 심포지엄
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• pp.40-64
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• 2005

Much biochemical information on peroxiredoxins (Prxs) has been reported but a genuine physiological function for these proteins has not been established. We show here that two cytosolic yeast Prxs, cPrxI and II, exist in a variety of forms that differ in their structure and molecular weight (MW) and that they can act both as a peroxidase and as a molecular chaperone. The peroxidase function predominates in the lower MW proteins, whereas the chaperone function is more significant in the higher MW complexes. Oxidative stress and heat shock exposure of yeasts causesthe protein structures of cPrxI and II to shift from low MW species to high MW complexes. This triggers a peroxidase-to-chaperone functional switch. These in vivo changes are primarily guided by the active peroxidase site residue, $Cys^{47}$, which serves as an efficient $'H_2O_2-sensor'$ in the cells. The chaperone function of the proteins enhances yeast resistance to heat shock.