國民醫療와 宗敎病院의 役割

柳寅旺 한국가톨릭병원협회 1975 한국가톨릭병원협회지 Vol.6 No.1

개업가-앞으로 어떻게 변할것인가

류인왕 釜山醫師會 1981 釜山醫師會誌 Vol.17 No.5

Thermoresistant properties of bacterioferritin comigratory protein against high temperature stress in Schizosaccharomyces pombe

류인왕,이수희,임혜원,안기섭,박광학,사재훈,정경진,임창진,김경훈,Ryu, In Wang,Lee, Su Hee,Lim, Hye-Won,Ahn, Kisup,Park, Kwanghark,Sa, Jae-Hoon,Jeong, Kyung Jin,Lim, Chang-Jin,Kim, Kyunghoon The Microbiological Society of Korea 2016 미생물학회지 Vol.52 No.4

The Schizosaccharomyces pombe structural gene encoding bacterioferritin comigratory protein (BCP) was previously cloned using the shuttle vector pRS316 to generate the BCP-overexpressing plasmid pBCP10. The present work aimed to evaluate the thermoresistant properties of BCP against high temperature stress using the plasmid pBCP10. When the S. pombe cells were grown to the early exponential phase and shifted from $30^{\circ}C$ to $37^{\circ}C$ or $42^{\circ}C$, the S. pombe cells harboring pBCP10 grew significantly more at both $37^{\circ}C$ and $42^{\circ}C$ than the vector control cells. After 6 h of the shifting to higher incubation temperatures, they contained the lower reactive oxygen species (ROS) and nitrite content, an index of nitric oxide (NO), than the vector control cells. After the temperature shifts, total glutathione (GSH) content and total superoxide dismutase (SOD) activities were much higher in the S. pombe cells harboring pBCP10 than in the corresponding vector control cells. Taken together, the S. pombe BCP plays a thermoresistant role which might be based upon its ability both to down-regulate ROS and NO levels and to up-regulate antioxidant components, such as total GSH and SOD, and subsequently to maintain thermal stability. 이전의 연구에서, bacterioferritin comigratory protein (BCP)을 인코딩하는 Schizosaccharomyces pombe의 구조유전자를 shuttle vector인 pRS316에 클로닝하여 BCP 과잉발현 플라즈미드인 pBCP10을 제조한 바 있다. 본 연구에서는, 플라즈미드 pBCP10을 사용하여 고온 스트레스에 대한 BCP의 열저항적 성질을 평가하였다. 대수기의 초기까지 성장시킨 S. pombe 세포의 배양 온도를 $30^{\circ}C$에서 $37^{\circ}C$나 $42^{\circ}C$로 전이시키는 경우, pBCP10 함유 S. pombe 세포가 벡터 대조 세포보다 $37^{\circ}C$와 $42^{\circ}C$ 모두에서 유의하기 더 잘 성장하였다. 높은 배양 온도로 전이한 뒤 6시간에서, pBCP10 함유 S. pombe 세포가 벡터 대조 세포보다 낮은 활성산소종(ROS)과 일산화질소(NO)의 지표로 측정된 아질산염(nitrite) 함량을 갖고 있음이 확인되었다. 온도 전이 뒤에, 총 글루타치온(total glutathione) 함량과 총 수퍼옥사이드 디스뮤타제(superoxide dismutase) 활성은 대응되는 벡터 대조 세포보다 pBCP10 함유 S. pombe 세포에서 현저하게 높다는 사실도 확인되었다. 종합하면, S. pombe BCP는 열저항적 역할을 보유하는 데, 활성산소종과 일산화질소에 대한 하강시키는 활성과 총 글루타치온과 수퍼옥사이드 디스뮤타제 등의 항산화 성분들을 상승시키는 활성, 즉 종합적으로 열안정성을 유지하는 활성에 근거하는 것으로 추정되었다

Molecular Cloning and Expression of a Laccase from Ganoderma lucidum, and Its Antioxidative Properties

주성수,류인왕,Ji-Kook Park,Yeong Min Yoo,Dong-Hyun Lee,Kwang Woo Hwang,최형태,임창진,이도익,김경훈 한국분자세포생물학회 2008 Molecules and cells Vol.25 No.1

Laccases are multicopper-containing oxidases that catalyze the oxidation of many aromatic compounds with concomitant reduction of oxygen to water. Interest in this enzyme has arisen in many fields of industry, including detoxification, wine stabilization, paper processing, and enzymatic conversion of chemical intermediates. In this study, we cloned a laccase gene (GLlac1) from the white-rot fungus Ganoderma lucidum. The cloned gene consists of 4,357 bp, with its coding region interrupted by nine introns, and the upstream region has putative CAAT and TATA boxes as well as several metal responsive elements (MREs). We also cloned a full-length cDNA of GLlac1, which contains an uninterrupted open reading frame (ORF) of 1,560 bp coding for 520 amino acids with a putative 21-residue signal sequence. The DNA and deduced amino acid sequences of GLlac1 were similar but not identical to those of other fungal laccases. GLlac1 was released from the cells when expressed in P. pastoris, and had high laccase activity. In addition, GLlac1 conferred antioxidative protection from protein degradation, and thus may be useful in bio-medical applications.

Defensive roles of Sdu1, a PPPDE superfamily member with ubiquitin C-terminal hydrolase activity, against thermal stress in Schizosaccharomyces pombe

한희,허태영,류인왕,김경훈,임창진,Han, Hee,Heo, Tae Young,Ryu, In Wang,Kim, Kyunghoon,Lim, Chang-Jin The Microbiological Society of Korea 2015 미생물학회지 Vol.51 No.4

Schizosaccharomyces pombe $sdu1^+$ 유전자는 PPPDE superfamily member에 속하는 탈유비퀴틴화 효소인 Sdu1을 엔코딩한다. 이전 연구에서, $sdu1^+$ 유전자 포함 재조합 플라스미드 pYSTP를 사용하여 Sdu1이 카르복시 말단 유비퀴틴 가수분해 활성을 보유한다는 사실이 입증된 바 있다. 본 연구에서는, 높은 배양 온도에 대한 Sdu1의 열내성적 역할이 검토되었다. 온도 천이 실험에서, pYSTP 포함 S. pombe 세포들이 37도나 42도로 천이한 후에 벡터 대조 세포들보다 훨씬 더 잘 성장하였다. 37도나 42도로 천이 한 후 6시간 배양한 pYSTP 포함 S. pombe 세포들이 벡터 대조 세포들보다 낮은 활성산소종 수준을 나타내었다. pYSTP 포함 S. pombe 세포들이 온도 천이와 관계없이 벡터 대조 세포들보다 다소 낮은 일산화질소 수준을 나타내었다. pYSTP 포함 S. pombe 세포들이 벡터 대조 세포들보다 훨씬 높은 총 글루타치온 수준을 나타내었다. 온도천이 후의 총 수퍼옥시드 디스무타아제 및 글루타치온 과산화효소 활성들이 pYSTP 포함 S. pombe 세포들에서 더 높은 것으로 측정되었다. 요약하면, S. pombe Sdu1은 활성산소종과 일산화질소 수준은 낮추고, 총 글루타치온, 총 수퍼옥시드 디스무타아제 및 글루타치온 과산화효소 수준들은 증가시킴으로써 높은 배양 온도에 대한 열내성적 역할을 나타낸다. The $sdu1^+$ gene encodes Sdu1, a PPPDE superfamily member of deubiquitinating enzymes (DUBs) in Schizosaccharomyces pombe. Sdu1 was previously shown to contain an actual ubiquitin C-terminal hydrolase (UCH) activity using the recombinant plasmid pYSTP which harbors the $sdu1^+$ gene. This work was designed to assess a thermotolerant role of Sdu1 against high incubation temperatures. In the temperature-shift experiments, the S. pombe cells harboring pYSTP grew much better after the shifts to $37^{\circ}C$ and $42^{\circ}C$, when compared with the vector control cells. After being shifted to $37^{\circ}C$ and $42^{\circ}C$ for 6 h, the S. pombe cells harboring pYSTP contained lower reactive oxygen species (ROS) levels, compared with the vector control cells. The nitric oxide (NO) levels of the S. pombe cells harboring pYSTP were slightly lower than those of the vector control cells in the absence or presence of the temperature shifting. The total glutathione (GSH) levels of the S. pombe cells harboring pYSTP were significantly higher than those of the vector control cells. Total superoxide dismutase (SOD) and GSH peroxidase activities were also higher in the S. pombe cells harboring pYSTP after the temperature shifts than in the vector control cells. In brief, the S. pombe Sdu1 plays a thermotolerant role against high incubation temperature through the down-regulation of ROS and NO and the up-regulation of total GSH content, total SOD and GSH peroxidase activities.

Carbon Source-Dependent Regulation of the Schizosaccharomyces pombe pbh1 Gene

Su-Jung Kim,Nam-Chul Cho,류인왕,김경훈,박은희,임창진 한국미생물학회 2006 The journal of microbiology Vol.44 No.6

Pbh1, from the fission yeast Schizosaccharomyces pombe, is a baculoviral inhibitor of apoptosis (IAP) repeat (BIR) domain-containing protein. Its unique encoding gene was previously found to be regulated by nitric oxide and nitrogen starvation. In the current work, the Pbh1-lacZ fusion gene was used to elucidate the transcriptional regulation of the pbh1 gene under various carbon sources. When fermentable carbon sources, such as glucose (at a low concentration of 0.2%), sucrose (2.0%) and lactose (2.0%), were the sole carbon source, the synthesis of β-galactosidase from the Pbh1-lacZ fusion gene was reasonably enhanced. However, the induction by these fermentable carbon sources was abolished in the Pap1-negative S. pombe cells, implying that this type of induction of the pbh1 gene is mediated by Pap1. Ethanol (2.0%), a nonfermentable carbon source, was also able to enhance the synthesis of β-galactosidase from the fusion gene in wild-type cells but not in Pap1-negative cells. The results indicate that the S. pombe pbh1 gene is up-regulated under metabolic oxidative stress in a Pap1-dependent manner.

Reactive oxygen species-dependent transcriptional regulation of peroxisome proliferator-activated receptor γ coactivator 1α in a human hepatocarcinoma cell line

김경훈,김수정,조남철,정현주,안기섭,류인왕,임창진 한국유전학회 2012 Genes & Genomics Vol.34 No.6

The present work aimed to examine whether transcriptional regulation of the human PGC-1α gene encoding peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α) is subjected to oxidative stress or not. Oxidative stressors, such as superoxide radical-generating menadione (MD) and hydrogen peroxide (H2O2), brought about a diminishment in the PGC-1α mRNA levels of the human hepatocarcinoma HepG2cells. The PGC-1α mRNA level was enhanced by the treatment with exogenous antioxidants, such as glutathione (GSH) and N-acetylcysteine (NAC). The HepG2 stable transfectants overexpressing the mitochondrial isoform of human glutaredoxin,Grx2a, were assessed to contain the significantly increased PGC-1α mRNA level, compared to the vector control cells. Taken together, transcription of the human PGC-1α gene is negatively regulated by reactive oxygen species (ROS) in a human hepatocarcinoma cell line.

생쥐 뇌소교세포주에서 웅담추출활성성분(우르소데옥시콜린산)의 항산화 및 세포보호효과

주성수(Seong-Soo Joo),김성근(Seong-Kun Kim),유영민(Yeong-Min Yoo),류인왕(In-Wang Ryu),김경훈(Kyunghoon Kim),이도익(Do-Ik Lee) 한국식품과학회 2006 한국식품과학회지 Vol.38 No.3