• 한국동물학회:학술대회논문집
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• 한국동물학회 1995년도 한국생물과학협회 학술발표대회
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• pp.248.2-248
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• 1995

No Abstract, See Full Text

• 한국결정학회:학술대회논문집
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• 한국결정학회 2002년도 정기총회 및 추계학술연구발표회
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• pp.25-25
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• 2002

CodWX, encoded by the cod operon in Bacillus subtilis, is a member of the ATP-dependent protease complex family, and is homologous to the eukaryotic 26S proteasome. It consists of two multimeric complexes: two hexameric ATPase caps of CodX and a protease chamber consisting of CodW dodecamer. Prior structural studies have shown that the N-terminal threonine residue is solely functional as a proteolytic nucleophile in ATP-dependent proteases such as HslV and certain β-type subunits of 20S proteasome, which have a primary sequence similarity of -50% and -20% with CodW respectively. Here we present a 3.0 Å resolution crystal structure of CodW, which is the first N-terminal serine protease among the known proteolytic enzymes. In spite of the same fold and the conserved contacts between subunits with HslV in E. coli and H. influenza, this structure shows the five additional residues extending from conserved Thr1 among the other ATP-dependent pretense and extraordinary basic proteolytic chamber.

• Plant Biotechnology Reports
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• 제3권3호
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• pp.225-241
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• 2009

We investigated the transcriptional profiles of Japanese indigenous grape cultivar 'Koshu' (Vitis vinifera) leaf and berry skin during ripening. In leaf, 64 genes were abundantly transcribed at the end of $v{\acute{e}}raison$ (14 weeks post-flowering), whereas the expression of 61 genes was upregulated at the end of ripening (20 weeks post-flowering). In berry skin, 67 genes were abundantly transcribed at the end of $v{\acute{e}}raison$, whereas the expression of 86 genes was upregulated at the end of ripening. Gene expression associated with biological processes was activated in both tissues at the end of ripening. The expression of genes associated with photosynthesis, sugar synthesis, anthocyanin synthesis, cinnamic acid synthesis, and amino acid metabolism was observed in leaf and berry skin during ripening, together with the accumulation of sugars, anthocyanins, cinnamic acids, and amino acids. Transcripts of AUX/IAA family proteins that repress the activities of auxin-induced proteins were expressed in berry skin at the end of $v{\acute{e}}raison$. Transcripts of genes related to the ubiquitin-proteasome system that degrades AUX/IAA family proteins were abundantly expressed in berry skin at the end of ripening, suggesting that the expansion of skin cells at $v{\acute{e}}raison$ is suppressed by AUX/IAA family proteins, and that the ubiquitin-proteasome system induces the expansion of skin cells during ripening by degrading AUX/IAA family proteins. These transcriptional profiles, which provide new information on the characteristics of 'Koshu' grapevine during ripening, may explain the unique characteristics of 'Koshu' grape in comparison with those of European grapes used for winemaking, and may contribute to the improvement of 'Koshu' grape quality.

• Journal of Photoscience
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• 제11권3호
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• pp.121-127
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• 2004

The${\beta}$-glucosidases occur widely in all living organisms and has in general a tendency to form oligomers of varying numbers of subunits or aggregates, although the functional implications of such diverse oligomerization schemes remain unclear. In particular, the assembly mode of the oat ${\beta}$-glucosidase is very unique in that it multimerizes by linear stacking of a hexameric building block to form long fibrillar multimers. Some structural proteins such as actin and tubulin assemble into long fibrils in a helical fashion and several enzymes such as GroEL and Pyrodictium ATPase functional complexes, 20S proteasome of the archaebacterium Thermoplasma acidophilum, and lutamine synthetase fromblue-green algae, assemble into discrete oligomers upto 4 stacked rings to maintain their enzymatic activities. In particular, oat ${\beta}$-glucosidase exists in vivo as a discrete long fibrillar multimer assembly that is a novel structure for enzyme protein. It is assembled by linear stacking of hollow trimeric units. The fibril has a long central tunnel connecting to the outer medium via regularly distributed side fenestrations. The enzyme active sites are located within the central tunnel and multimerization increases enzyme affinity to the substrates and catalytic efficiency of the enzyme. Although it is suggested that oligomerization may contribute to the enzyme stability and catalytic efficiency of ${\beta}$-glycosidases, the functional implications of such diverse oligomerization schemes remain unclear so far.

• 한국초지조사료학회지
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• 제25권4호
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• pp.287-296
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• 2005

프로테오믹스 기법을 이용하여 벼 저온 스트레스 관련 단백질을 분리 동정하기 위하여 저온 처리한 벼로부터 단백질을 분리하였다. 분리한 단백질로부터 Rubisco 단백질을 제거하기 위해 $15\%$ PEG fractionation을 실시한 후 $15\%$ PEG 상등액과 pellet 분획을 각각 이차원전기 영동으로 단백질을 분석하였고, MALDI-TOF MS를 이용하여 단백질을 동정하였다. $15\%$ PEG 상등액에서 8개의 단백질 spot이 증가하였고 10개의 spot 이 감소하였다. 증가한 8개 단백질 spot 중에서 epimerase/dehydratase, fructokinase, ribose-5-phosphate isomerase (Rpi), chaperonin 21 precursor, photosystem II oxygen-envolving complex (PS II OEC) protien 2 precursor, thioredoxin h-type (Trx-h) 등 6개의 단백질이 확인되어졌다. $15\%$ PEG pellet 분획에서 13개의 단백질 spot이 증가하였고 14 spot이 감소하였으며, 증가한 13개 단백질 spot중에서 OSJNB b059K02.15, hypothetical protein, mitogen-activated protein kinase kinase (MAPKK), 20S proteasome beta 7 subunit, Rubisco small subunit 등 5개의 단백질이 확인되어졌다. 확인되어진 단백질들은 기능별로 분류해 본 결과, 세포대사관련 단백질, energy 생성에 관련된 단백질, 산화환원 조절관련 단백질, 식물 병 방어관련, 단백질 합성 및 신호전달 관련 단백질 등으로 분류되었다. 이들 중 RPi와 MAPKK가 저온 스트레스에 의해 발현되는 것이 본 실험의 프로테옴 분석을 통하여 최초로 동정되었다.

• Asian-Australasian Journal of Animal Sciences
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• 제22권1호
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• pp.20-25
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• 2009

Xenotransplantation of pig organs into humans is a potential solution for the shortage of donor organs for transplantation. However, multiple immune barriers preclude its clinical application. In particular, the initial type of rejection in xenotransplantation is an acute cellular rejection by host $CD8^+$ cytotoxic T lymphocyte (CTL) cells that react to donor major histocompatibility complex (MHC) class I. The human cytomegalovirus (HCMV) glycoprotein Unique Short (US) 2 specifically targets MHC class I heavy chains to relocate them from the endoplasmic reticulum (ER) membrane to the cytosol, where they are degraded by the proteasome. In this study we transfected the US2 gene into minipig fetal fibroblasts and established four US2 clonal cell lines. The integration of US2 into transgenic fetal cells was confirmed using PCR and Southern blot assay. The reduction of Swine Leukocyte Antigen (SLA)-I by US2 was also detected using Flow cytometry assay (FACS). The FACS analysis of the US2 clonal cell lines demonstrated a substantial reduction in SLA-I surface expression. The level (44% to 76%) of SLA-I expression in US2 clonal cell lines was decreased relative to the control. In cytotoxicity assay the rate of $CD8^+$ T cell-mediated cytotoxicity was significantly reduced to 23.8${\pm}$15.1% compared to the control (59.8${\pm}$8.4%, p<0.05). In conclusion, US2 can directly protect against $CD8^+$-mediated cell lysis. These results indicate that the expression of US2 in pig cells may provide a new approach to overcome the CTL-mediated immune rejection in xenotransplantation.

• Tuberculosis and Respiratory Diseases
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• 제48권2호
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• pp.166-179
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• 2000

연구배경: 현재 사용되고 있는 많은 항암제는 암세포의 apoptosis를 유도함으로서 암세포를 사멸시키는 것이 주된 작용 기전이다. 항암제에 의한 치료 실패의 주된 원인은 암세포의 apoptosis에 대한 내성 획득에 의한 것으로 이해되고 있다. 최근의 보고에 의하면 많은 암세포주에서 apoptosis에 대한 내성 획득에 NF-${\kappa}B$의 활성화가 중요한 역할을 한다고 알려져 있지만 일부 세포에서는 상반된 결과를 보이고 있어 apoptosis에 대한 내성 획득과 NF-${\kappa}B$의 활성화와의 관련성은 세포에 따라 차이를 보이고 있다. 본 연구에서는 비소세포폐암 세포주가 TNF-$\alpha$ 유발 apoptosis에 저항을 나타내는 기전에서 NF-${\kappa}B$의 역할을 규명하고자 하였다. 방 법: 비소세포폐암세포주인 NCI-H157 세포에 TNF-$\alpha$ cycloheximide(CHX), TNF-$\alpha$와 CHX를 각각 투여하고 24시간 후 MTT assay로 세포생존율을 평가하였고 apoptosis의 발생 유무는 PARP에 대한 Westrn 분석으로 평가하였다. Ad5LacZ와 $Ad5I{\kappa}B{\alpha}SR$를 20시간 동안 감염시킨 각각의 세포를 1, 5, 10, 20, 50 ng/ml의 TNF-$\alpha$로 24, 48시간 자극 후 MTT assay를 시행하였고, 같은 농도의 TNF-$\alpha$로 24, 48시간 자극 후 PARP에 대한 Western 분석을 시행하였다. $I{\kappa}B{\alpha}$의 분해를 억제하는 proteasome inhibitor 인 MG132를 전처치하고 TNF-$\alpha$로 24, 48 시간 자극 후 MTT assay와 PARP에 대한 Westrn 분석을 시행하였다. $Ad5I{\kappa}B{\alpha}SR$를 감염시킨 세포를 TNF-$\alpha$로 자극한 후 NF-${\kappa}B$의 활성화를 EMSA로 평가하였다. 결 과: 1. TNF-$\alpha$ 단독 투여로는 세포 생존율의 감소와 apoptosis가 관찰되지 않았고 TNF-$\alpha$와 CHX를 같이 투여하였을 때는 세포 생존율의 감소와 함께 apoptosis가 유도되었다. 2. $Ad5I{\kappa}B{\alpha}SR$의 감염으로 $I{\kappa}B{\alpha}$가 과발현된 세포와 MG132를 전처치한 세포에서 TNF-$\alpha$ 자극에 의한 NF-${\kappa}B$의 활성화가 억제되었다. 3. $Ad5I{\kappa}B{\alpha}$ SR를 감염시킨 세포와 MG132를 전처치하여 NF-${\kappa}B$의 활성을 억제한 세포에서 TNF-$\alpha$ 자극 후 세포 생존율이 감소하고 apoptosis가 유도되었다. 결 론: 비소세포폐암 세포주의 TNF-$\alpha$ 유발 apoptosis에 대한 내성은 NF-${\kappa}B$의 활성화에 의해 생성되는 새로운 단백의 발현에 의한 것으로 생각되며, NF-${\kappa}B$ 활성화의 억제로 이를 극복할 수 있을 것으로 사료된다.

• 한국축산식품학회지
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• 제42권2호
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• pp.266-279
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• 2022

This study was conducted to evaluate the proteolysis trends and change in meat quality during 10 days of cold storage in duck M. pectoralis major (PM) and M. iliotibialis (IL). Duck IL had a higher pH and greater degree of lightness but lower cooking loss than PM (p<0.05). During the 10-day cold storage, the pH value of PM declined significantly (p<0.05), while the meat quality traits of IL were not affected by cold storage (p>0.05). In PM, the redness increased from day 1 to day 5, while cooking loss was lower on day 10 compared to day 5 (p<0.05). There were no significant differences in the activities of cathepsin B and proteasome 20S during cold storage (p>0.05). The activity of calpains declined gradually during 10 days of storage (p<0.05), and the activity of calpains in PM was higher than that in IL (p<0.05). A total of 5,155 peptides were detected and derived from 34 proteins of duck PM muscle, whereas 4,222 peptides derived from 32 proteins were detected from duck IL muscle. Duck PM muscle was composed only of fast type of muscle fiber, whereas IL muscle was composed of both slow and fast types. The proteins responsible for glycolysis or myofibrillar proteins were closely related to changes in meat color or water-holding capacity during cold storage. These results suggest that changes in meat quality characteristics during cold storage are closely related to protein degradation, which is also related to the distribution of muscle fiber types.