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Expression and Characterization of Truncated Recombinant Human Cytochrome P450 2J2
Hyoung-Goo Park,Young-Ran Lim,Songhee Han,Donghak Kim 한국독성학회 2014 Toxicological Research Vol.30 No.1
The human cytochrome P450 2J2 catalyzes an epoxygenase reaction to oxidize various fatty acids including arachidonic acid. In this study, three recombinant enzyme constructs of P450 2J2 were heterologously expressed in Escherichia coli and their P450 proteins were successfully purified using a Ni<SUP>2+</SUP>-NTA affinity column. Deletion of 34 amino acid residues in N-terminus of P450 2J2 enzyme (2J2-D) produced the soluble enzyme located in the cytosolic fraction. The enzymatic analysis of this truncated protein indicated the typical spectral characteristics and functional properties of P450 2J2 enzyme. P450 2J2-D enzymes from soluble fraction catalyzed the oxidation reaction of terfenadine to the hydroxylated product. However, P450 2J2-D enzymes from membrane fraction did not support the P450 oxidation reaction although it displayed the characteristic CO-binding spectrum of P450. Our finding of these features in the N-terminal modified P450 2J2 enzyme could help understand the biological functions and the metabolic roles of P450 2J2 enzyme and make the crystallographic analysis of the P450 2J2 structure feasible for future studies.
( Hyoung-goo Park ),( Young-ran Lim ),( Songhee Han ),( Dabin Jeong ),( Donghak Kim ) 한국미생물 · 생명공학회 2017 Journal of microbiology and biotechnology Vol.27 No.5
NADPH-P450 reductase (NPR) transfers electrons from NADPH to cytochrome P450 and heme oxygenase enzymes to support their catalytic activities. This protein is localized within the endoplasmic reticulum membrane and utilizes FMN, FAD, and NADPH as cofactors. Although NPR is essential toward enabling the biochemical and pharmacological analyses of P450 enzymes, its production as a recombinant purified protein requires a series of tedious efforts and a high cost due to the use of NADP+ in the affinity chromatography process. In the present study, the rat NPR clone containing a 6× Histidine-tag (NPR-His) was constructed and heterologously expressed. The NPR-His protein was purified using Ni<sup>2+</sup>-affinity chromatography, and its functional features were characterized. A single band at 78 kDa was observed from SDS-PAGE and the purified protein displayed a maximum absorbance at 455 nm, indicating the presence of an oxidized flavin cofactor. Cytochrome c and nitroblue tetrazolium were reduced by purified NPR-His in an NADPH-dependent manner. The purified NPR-His successfully supported the catalytic activities of human P450 1A2 and 2A6 and fungal CYP52A21, yielding results similar to those obtained using conventional purified rat reductase. This study will facilitate the use of recombinant NPR-His protein in the various fields of P450 research.
Expression and Characterization of Truncated Recombinant Human Cytochrome P450 2J2
Park, Hyoung-Goo,Lim, Young-Ran,Han, Songhee,Kim, Donghak Korean Society of ToxicologyKorea Environmental Mu 2014 Toxicological Research Vol.30 No.1
The human cytochrome P450 2J2 catalyzes an epoxygenase reaction to oxidize various fatty acids including arachidonic acid. In this study, three recombinant enzyme constructs of P450 2J2 were heterologously expressed in Escherichia coli and their P450 proteins were successfully purified using a $Ni^{2+}$-NTA affinity column. Deletion of 34 amino acid residues in N-terminus of P450 2J2 enzyme (2J2-D) produced the soluble enzyme located in the cytosolic fraction. The enzymatic analysis of this truncated protein indicated the typical spectral characteristics and functional properties of P450 2J2 enzyme. P450 2J2-D enzymes from soluble fraction catalyzed the oxidation reaction of terfenadine to the hydroxylated product. However, P450 2J2-D enzymes from membrane fraction did not support the P450 oxidation reaction although it displayed the characteristic CO-binding spectrum of P450. Our finding of these features in the N-terminal modified P450 2J2 enzyme could help understand the biological functions and the metabolic roles of P450 2J2 enzyme and make the crystallographic analysis of the P450 2J2 structure feasible for future studies.
Park, See-Hyoung,Lee, Jongsung,Shon, Jong Cheol,Phuc, Nguyen Minh,Jee, Jun Goo,Liu, Kwang-Hyeon Elsevier 2018 Phytomedicine Vol.42 No.-
<P>Conclusion: Overall, this was the first report of the inhibitory effects of BCA on CYP2J2 in HLMs. The present data suggest that BCA is a potential candidate for further evaluation for its CYP2J2 targeting anti-cancer activities.</P>
교합성 골수강내 금속정과 금속판 내고정술을 이용한 상완골 간부골절 치료의 비교
박홍기,이수찬,김형일,강기동,고진홍,문도현,이범구 대한골절학회 1997 대한골절학회지 Vol.10 No.3
Most of the humeral shaft fractures have been treated conservatively. If operation is indicated, most surgeons used to perform an open reduction and internal fixation with plate & screws. But, when open reduction and internal fixation is carried out, the periosteum and soft tissue attachments must be stripped off from the bone and operative time becomes longer. Interlocking IM nailing is an attractive procedure which does not disrupt or strip off periosteum and soft tissue at the fracture site, decrease the chance of infection and allow early exercise. Authors compare the clinical result of humerus shaft fracture which were treated by plate fixation in 25 patients and interlocking IM nailing in 16 patients who were treated at the orthopedic department, Joong Ang Gil Hospital from Feb. 1992 to Jan. 1995. 1. The average time of operation in interlocking IM nail was 65 minutes and that of plate fixation was 95 minutes. 2. The average time for bone union was about 14.4 weeks in interlocking IM nail and 14.2 weeks in plate fixation. 3. The complications of interlocking IM nail were 1 case of postoperative radial nerve palsy, 1 case of delayed union, 1 case of nonunion and 3 cases of pain and stiffness of shoulder. 4. The complications of plate fixation were 1 case of infection, 1 case of delayed union, 3 cases of nonunion, 1 case of metal failure, 2 cases of postoperative radial nerve palsy and 2 cases of pain and stiffness of elbow. In conclusion, the interlocking IM nailing is an attractive procedure for the treatment of the humeral shaft fractures
박종민,김만수,권순구,엄순형 한국산업식품공학회 1998 산업 식품공학 Vol.2 No.2
제품의 포장에 있어 완충제의 역할은 제품의 유통과정중에 발생하는 충격과 진동을 완화시켜 제품을 보호하고 적정포장비로 유통합리화에 기여한다. 즉, 포장할 제품의 특성 및 물류환경에 따라 포장설계를 할 때 완충제의 선택 및 사용량 결정은 완충재 자체의 특성에 의존한다. 현재 보급되어 있는 완충시험가들은 비교적 얇은 재료에 대한 시험을 할 수 없고, 그 분석항목도 매우 제한적 이므로 본 연구에서는 박판재료로부터 후판재료까지 그 적용범위가 넓고, 재료의 완충성을 다각적으로 분석할 수 있도록 데이터와 측정부터 분석 및 출력까지 전 시스템을 일체화시킨 동적완충시험기를 설계하였다. 설계된 동적완충시험기의 성능시험에는 가속도 측정에 의해 계산된 동적응력 값과 실측에 의한 동적응력 값간의 차이분석과 동일조건에 대한 설계한 시험기와 기존의 시험기에 의한 시험결과의 분석 등의 방법을 적용하였는데, 두 경우 모두에서 비교적 잘 일치하는 결과를 얻었다. The most important thing for cushion packaging design of products is the physical properties of the cushioning, materials. For the cushioning performance analysis, the dynamic cushioning test, which is based on the real situations, is generally employed. This study was carried out to develop a low cost multi-purpose impact tester which can test a wide range of materials from thin to thick packaging materials, and can also analyze the shock-absorbing characteristics of materials in various aspects. The designed tester was verified by the differential analysis between measured dynamic stress and calculated dynamic stress and by the analysis of the results obtained from the existing tester. According to the aforementioned tests, results were relatively coincided with each other.
Peroxiredoxin 2 Deficiency Exacerbates Atherosclerosis in Apolipoprotein E-Deficient Mice
Park, Jong-Gil,Yoo, Ji-Young,Jeong, Se-Jin,Choi, Jae-Hoon,Lee, Mi-Ran,Lee, Mi-Ni,Hwa Lee, Jeong,Kim, Hyoung Chin,Jo, Hanjoong,Yu, Dae-Yeul,Kang, Sang Won,Rhee, Sue Goo,Lee, Mun-Han,Oh, Goo Taeg Ovid Technologies Wolters Kluwer -American Heart A 2011 Circulation research Vol.109 No.7
Peltier Heating-Assisted Low Temperature Plasma Ionization for Ambient Mass Spectrometry
( Hyoung Jun Lee ),( Ji-seon Oh ),( Sung Woo Heo ),( Jeong Hee Moon ),( Jeong-hoon Kim ),( Sung Goo Park ),( Byoung Chul Park ),( Gi Ryang Kweon ),( Yong-hyeon Yim ) 한국질량분석학회 2015 Mass spectrometry letters Vol.6 No.3
Low temperature plasma (LTP) ionization mass spectrometry (MS) is one of the widely used ambient analysis methods which allows soft-ionization and rapid analysis of samples in ambient condition with minimal or no sample preparation. One of the major advantages of LTP MS is selective analysis of low-molecular weight, volatile and low- to medium-polarity analytes in a sample. On the contrary, the selectivity for particular class of compound also implies its limitation in general analysis. One of the critical factors limiting LTP ionization efficiency is poor desorption of analytes with low volatility. In this study, a home-built LTP ionization source with Peltier heating sample stage was constructed to enhance desorption and ionization efficiencies of analytes in a sample and its performance was evaluated using standard mixture containing fatty acid ethyl esters (FAEEs). It was also used to reproduce the previous bacterial identification experiment using pattern-recognition for FAEEs. Our result indicates, however, that the bacterial differentiation from FAEE pattern recognition using LTP ionization MS still has many limitations.