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Biochemical analysis of recombinant CYP4A11 allelic variant enzymes: W126R, K276T and S353G
Han, S.,Cha, G.S.,Chun, Y.J.,Lee, C.H.,Kim, D.,Yun, C.H. JAPANESE SOCIETY FOR THE STUDY OF XENOBIOTICS 2016 DRUG METABOLISM AND PHARMACOKINETICS Vol.31 No.6
Human CYP4A11 is the major ω-hydroxylase of fatty acids in the liver and kidneys. It produces 20-hydroxyeicosatetraenoic acid as well as hydroxylates fatty acids. In this study, we investigated the biochemical properties of three alleles of CYP4A11: W126R, K276T, and S353G. Site-directed mutagenesis of the wild type CYP4A11 was performed, to construct the W126R, K276T, and S353G variant clones. The CYP4A11 wild type and variant constructs were heterologously expressed in Escherichia coli. CO-binding spectra showed the expression of the wild type, K276T and S353G variants, indicating the functional P450 holoenzyme. The W126R variant was not expressed in E. coli. Binding affinities of lauric acid in K276T and S353G variants were stronger than that of wild type. Steady-state kinetics in the hydroxylation reaction of fatty acids were studied. The catalytic efficiencies (k<SUB>cat</SUB>/K<SUB>m</SUB>) of K276T and S353G variants in the reactions without cytochrome b<SUB>5</SUB> were approximately 2- and 4-fold higher, respectively, than that of wild type, and in the reactions with cytochrome b<SUB>5</SUB> they were approximately 2- and 3-fold higher, respectively. These results suggest that individuals carrying the alleles, K276T and S353G, might exhibit higher catalysis of CYP4A11, which may affect the endogenous metabolic products associated with regulation of blood pressure.
비만에 따른 요통환자의 체중분포와 요부 근력차이에 관한 비교분석
한길수 ( G. S. Han ),김건도 ( G. D. Kim ),임동춘 ( D. C. Lim ) 한국운동생리학회(구-한국운동과학회) 2010 운동과학 Vol.19 No.4
이 연구의 목적은 비만에 따른 요통환자의 체중분포와 요부 근기능 및 안정화 비율의 차이를 비교·분석하는 데 있다. 서울 강남에 소재한 J병원에 내원한 남성 만성요통환자 60명 중 체질량지수(Body Mass Index: BMI)가 25kg/m² 이상인 G1그룹 30명과 이하인 G2그룹 30명을 대상으로 Tetrax와 MedX를 이용하여 분석한 결과 다음과 같은 결론을 얻었다. 체중분포에서 G1그룹은 왼쪽 발뒤꿈치 1.97%(p>.05), 오른쪽 발뒤꿈치 2.37%에서 높게 나타났고(p>.05), G2그룹에서는 왼쪽 발앞꿈치 2.75%(p>.05), 오른쪽 발앞꿈치 1.97%(p>.05)에서 더 높은 체중이 실리는 것으로 나타났다. 요부 각도별 신전근력에서 G1그룹이 G2그룹에 비해 모든 각도에서 높은 근력을 나타냈고, 0°(p>.05), 12°(p>.05)를 제외한 24°(p<.05), 36°(p<.01), 48°(p<.001), 60°(p<.001), 72°(p<.001)에서 통계적으로 유의한 차이를 나타냈다. 요부 안정화 비율에서는 G1그룹의 경우 2.75+_1.31, G2그룹 2.48+_2.23로 나타났다(p>.05). 결론적으로 요부 각도별 신전근력에서 G1그룹이 G2그룹에 비해 모든 각도에서 근력이 높게 나타냈고. 발의 체중분포에서 G1그룹은 비만에 따른 요추의 전만을 증가시켜 힘의 중심을 더 후방으로 유지하려는 경향을 보이고 있는데, 이는 요부 신전근력의 약화로 이어져 안정화 비율에도 영향을 미치는 것으로 사료된다. This study was aimed to determine the effect for the weight distribution and lumbar extension strength associated with obesity index in male patients of Chronic Low Back Pain. Sixty subjects(obesity group(BMI:25Kg/m2): n=30, non obesity group: n=30) participated on this study. Both group were tested on lumbar extension using by MedX machine. Front heels and back heels were measured twice using by the Tetrax Portable Multiple System. Independent sample t-test was used to analyze the difference between the two groups. Study results showed that the obese back pain group(G1) had more weight loaded on both heels than the normal back pain group(G2), but there was no significant difference between the groups (p>.05). As for the lumbar extension strength at each angle, the obese back pain group appeared higher muscular strength than the normal group at 0°, but was stronger at angles 12°, 24°, 36°, 48°, 60°, and 72°. The ratio of lumbar flexion 72° and 0° angle showed 2.75:1 in G1 group and 2.48:1 in G2 group. Overall, the back pain group of obesity(G1) according to weight distribution tended to maintain their center of force more backwards by increasing the lordosis degree with abdominal distension.
Han, J,Kim, Y-L,Lee, K-W,Her, N-G,Ha, T-K,Yoon, S,Jeong, S-I,Lee, J-H,Kang, M-J,Lee, M-G,Ryu, B-K,Baik, J-H,Chi, S-G Macmillan Publishers Limited 2013 Cell death and differentiation Vol.20 No.8
ZNF313 encoding a zinc-binding protein is located at chromosome 20q13.13, which exhibits a frequent genomic amplification in multiple human cancers. However, the biological function of ZNF313 remains largely undefined. Here we report that ZNF313 is an ubiquitin E3 ligase that has a critical role in the regulation of cell cycle progression, differentiation and senescence. In this study, ZNF313 is initially identified as a XIAP-associated factor 1 (XAF1)-interacting protein, which upregulates the stability and proapoptotic effect of XAF1. Intriguingly, we found that ZNF313 activates cell cycle progression and suppresses cellular senescence through the RING domain-mediated degradation of p21<SUP>WAF1</SUP>. ZNF313 ubiquitinates p21<SUP>WAF1</SUP> and also destabilizes p27<SUP>KIP1</SUP> and p57<SUP>KIP2</SUP>, three members of the CDK-interacting protein (CIP)/kinase inhibitor protein (KIP) family of cyclin-dependent kinase inhibitors, whereas it does not affect the stability of the inhibitor of CDK (INK4) family members, such as p16<SUP>INK4A</SUP> and p15<SUP>INK4B</SUP>. ZNF313 expression is tightly controlled during the cell cycle and its elevation at the late G1 phase is crucial for the G1-to-S phase transition. ZNF313 is induced by mitogenic growth factors and its blockade profoundly delays cell cycle progression and accelerates p21<SUP>WAF1</SUP>-mediated senescence. Both replicative and stress-induced senescence are accompanied with ZNF313 reduction. ZNF313 is downregulated during cellular differentiation process in vitro and in vivo, while it is commonly upregulated in many types of cancer cells. ZNF313 shows both the nuclear and cytoplasmic localization in epithelial cells of normal tissues, but exhibits an intense cytoplasmic distribution in carcinoma cells of tumor tissues. Collectively, ZNF313 is a novel E3 ligase for p21<SUP>WAF1</SUP>, whose alteration might be implicated in the pathogenesis of several human diseases, including cancers.
S6K1 Phosphorylation of H2B Mediates EZH2 Trimethylation of H3: A Determinant of Early Adipogenesis
Yi, S.,Um, S.,Lee, J.,Yoo, J.,Bang, S.,Park, E.,Lee, M.,Nam, K.,Jeon, Y.,Park, J.,You, J.,Lee, S.J.,Bae, G.U.,Rhie, J.,Kozma, Sara C.,Thomas, G.,Han, J.W. Cell Press 2016 Molecular Cell Vol.62 No.3
S6K1 has been implicated in a number of key metabolic responses, which contribute to obesity. Critical among these is the control of a transcriptional program required for the commitment of mesenchymal stem cells to the adipocytic lineage. However, in contrast to its role in the cytosol, the functions and targets of nuclear S6K1 are unknown. Here, we show that adipogenic stimuli trigger nuclear translocation of S6K1, leading to H2BS36 phosphorylation and recruitment of EZH2 to H3, which mediates H3K27 trimethylation. This blocks Wnt gene expression, inducing the upregulation of PPARγ and Cebpa and driving increased adipogenesis. Consistent with this finding, white adipose tissue from S6K1-deficient mice exhibits no detectable H2BS36 phosphorylation or H3K27 trimethylation, whereas both responses are highly elevated in obese humans or in mice fed a high-fat diet. These findings define an S6K1-dependent mechanism in early adipogenesis, contributing to the promotion of obesity.
Han, S.E.,Lee, H.G.,Yun, C.H.,Hong, Z.S.,Kim, S.H.,Kang, S.K.,Kim, S.H.,Cho, J.S.,Ha, S.H.,Choi, YunJaie Asian Australasian Association of Animal Productio 2005 Animal Bioscience Vol.18 No.12
Zinc is a trace element that is associated with a stimulation of immune function and regulation of ion balance for livestock production. In this study, the effect of zinc as inhibitor to apoptosis-induced cells was examined in vitro using mammary epithelial cell line, HC11 and macrophage cell line, NCTC3749. Cell viability, measured by MTT assay, indicated that 10 g/ml of zinc had a negative impact on cellular activity and 50 ng/ml was chosen for further testing. Apoptosis was induced in cells treated with C2-ceramide in serum-free media. DNA fragmentation and gene expression of acidic sphingomyelinase (a gene responsible for the progress of apoptosis) were distinctively low in zinc treated cells compared with those in non-treated controls. In conclusion, zinc is involved in the regulation of cell proliferation and apoptosis in mammary epithelial cells and macrophages.
Han, J.H.,Cha, M.J.,Kim, B.G.,Kim, S.K.,Min, K.S. Elsevier 2008 Inorganic chemistry communications Vol.11 No.7
Novel nickel(II) hexaaza macrocyclic complexes, [Ni(L<SUP>R,R</SUP>)](ClO<SUB>4</SUB>)<SUB>2</SUB> (1) and [Ni(L<SUP>S,S</SUP>)](ClO<SUB>4</SUB>)<SUB>2</SUB> (2), containing chiral pendant groups have been synthesized by an efficient one-pot template condensation and characterized (L<SUP>R,R/S,S</SUP>=1,8-di((R/S)-α-methylbenzyl)-1,3,6,8,10,13-hexaazacyclotetradecane). The crystal structures of 1 and 2 were determined by single-crystal X-ray analysis. Each complex has a square-planar coordination environment for the nickel(II) ion, and is either an R or an S enantiomorph depending on the pendant groups. The circular dichroism spectrum of 1 showed a negative, positive and negative peak at 345, 440, and 492nm, respectively, and that of 2 exhibited an enantiomeric pattern.
Han, J.E.,Lee, S.,Jeong, D.G.,Yoon, S.W.,Kim, D.J.,Lee, M.S.,Kim, H.K.,Park, S.K.,Kim, J.H.,Park, S.C. Published by Elsevier Ltd. on behalf of Internatio 2017 Journal of global antimicrobial resistance Vol.10 No.-
<P>Conclusions: To the best of our knowledge, this is the first complete genome of S. cohnii, and will provide important insights into the biodiversity of CoNS and valuable information for the control of this emerging pathogen. (C) 2017 International Society for Chemotherapy of Infection and Cancer. Published by Elsevier Ltd. All rights reserved.</P>
Hong, S.W.,Baik, J.E.,Kang, S.S.,Yun, C.H.,Seo, D.G.,Han, S.H. Pergamon Press 2014 Molecular immunology Vol.57 No.2
Streptococcus mutans is a pathogenic Gram-positive bacterium that is closely associated with dental caries and subsequent pulpal inflammation. Although lipoteichoic acid (LTA) is considered a major virulence factor of Gram-positive bacteria, little is known about the innate immunity to S. mutans LTA. In this study, we purified LTA from S. mutans (Sm.LTA) through n-butanol extraction, hydrophobic interaction column chromatography, and ion-exchange column chromatography to investigate its immunological properties using murine macrophages. The Sm.LTA preparation had no detectable contamination with endotoxins, proteins, or nucleic acids. Upon exposure to Sm.LTA, the murine macrophage cell-line RAW 264.7 cells produced TNF-α and nitric oxide (NO) in a dose-dependent manner. Sm.LTA preferentially bound to and activated CHO/CD14/TLR2 cells rather than CHO/CD14/TLR4 cells, which are stable transfectants expressing CD14 and TLR2 or CD14 and TLR4, respectively. Sm.LTA could not induce TNF-α or NO production in macrophages derived from TLR2-deficient mice whereas it dose-dependently induced those inflammatory mediators in wild-type macrophages. TLR2-dependent induction of NO by Sm.LTA was also confirmed in RAW 264.7 cells using specific antibodies blocking TLR2. Furthermore, Sm.LTA deacylated by alkaline hydrolysis neither stimulated TLR2 nor induced TNF-α or NO production, suggesting that Sm.LTA lipid moieties are crucial for the immuno-stimulatory activity of Sm.LTA. Unlike Staphylococcus aureus LTA, which has potent immuno-stimulating activity, Sm.LTA showed a modest induction of NO production comparable to LTAs of other oral bacteria Enterococcus faecalis and Lactobacillus plantarum. In conclusion, our results suggest that the Sm.LTA interacts with TLR2 through the lipid moiety for the induction of inflammatory mediators in macrophages.