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991.
Nawarak J Huang-Liu R Kao SH Liao HH Sinchaikul S Chen ST Cheng SL 《Journal of proteome research》2008,7(9):3737-3746
Although the toxicogenomics of kojic acid treated A375 human malignant melanoma cells has been elucidated, the proteomics of cellular response is still poorly understood. We performed proteomic analysis to investigate the anticancer effect of kojic acid on protein expression profile in A375 cells. A375 cells were treated with kojic acid at 8 microg/mL for 24, 48, and 72 h. With the use of 2-D PAGE and MALDI-Q-TOF MS and MS/MS analyses, proteomic profiles of A375 cells between control and kojic acid treatment were compared, and 30 differentially expressed proteins, containing 2 up-regulated proteins and 28 down-regulated proteins, were identified. Among these proteins, 17 isoforms of 5 identical proteins were observed and 11 chaperone proteins showed the high proportion of protein spots with 36.7% of total proteins. Bioinformatic tools were used to search for protein function and prediction of protein interaction. Sixteen differentially expressed proteins exhibited interaction network linked to the downstream regulations of p53 tumor suppressor and cell apoptosis, which may lead to suppress the melanogenesis and tumorigenesis of kojic acid treated A375 cells. In addition, GRP75, VIME and 2AAA were validated by Western blot analysis, whereas GRP75, 2AAA, HS90B, ENPL and KPYM were validated by RT-PCR. Therefore, these proteins play the important roles in cancer progression and may be potential biomarkers that are useful for diagnostic and therapeutic applications of malignant melanoma cancer. 相似文献
992.
谭翠燕 徐春和 沈均人 SAKUMA Shinsuke YAMAMOTO Yasushi BALNY Claude 阮康成 《Acta biochimica et biophysica Sinica》2003,(7)
利用荧光光谱学等方法结合高压力技术研究了光合作用系统II中的一个外周蛋白——— 2 3kD(以P2 3k表示 )蛋白的去折叠。热力学研究表明 ,在 2 0℃、180MPa(1MPa =10 .0大气压 )可使该蛋白质完全去折叠 ,而在3℃ ,16 0MPa即可使该蛋白质完全去折叠 ,这是迄今为止有关研究中最易被高压力去折叠的一个蛋白质。在2 0℃ ,该蛋白质在常压下去折叠反应的标准自由能与标准体积变化分别为 2 3.4 5kJ mol和 - 15 0 .3ml mol;动力学研究揭示该蛋白质的折叠反应的活化体积ΔV f 为正值 (84 .1ml mol) ,而去折叠反应的活化体积ΔV u 为负值(- 6 6 .2ml mol)。在常压下 ,折叠和去折叠反应的速度常数 (K0f,K0u)分别为 1.87s- 1 和 1.3× 10 - 4s- 1 ,这些结果为解释该蛋白质易被压力去折叠提供了线索 相似文献
993.
Alkaline phosphatase(AKP),from the succus entericus of silkworm,was purified using 10%-50% ammonium sulfate fractions,ion exchange chromatography Of DEAE-Sepharose,and size exclusion chromatography of Sephacryl S-200.The purification fold was 464 times and specified activity was 3936 U/mg.Optimum pH value of the phosphatase was 10.5,and was stable between pH 7.5 and 11.The optimum temperature of the phosphatase was 40℃ and it was unstable over 50℃.Km value of the phosphatase was 1.25 mmol/L.In a given condition,the phosphatase was selectively modified by PCMB,NBS,PMSE TNBS,SUAN,DTT,BrAc,and IAc,the results indicate that PMSF,SUA,BrAc,IAc,and TNBS could Obviously inhibit the activity of the phosphatase,and the degree of inhibition depended on the concentration of these reagents.There was little effect on the activity of phosphatase after treatment by PMSF,DTT,and NBT.We primarily conclude that mercapto and imidazole are essential for AKP from silkworm.Also,Lys residue and disulfide bands are necessary to protect the catalysis of the AKP. 相似文献
994.
Increased levels of Wee-1 kinase in G(2) are necessary for Vpr- and gamma irradiation-induced G(2) arrest
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Human immunodeficiency virus type 1 (HIV-1) Vpr induces cell cycle arrest at the G(2)/M transition and subsequently apoptosis. Here we examined the potential involvement of Wee-1 in Vpr-induced G(2) arrest. Wee-1 is a cellular protein kinase that inhibits Cdc2 activity, thereby preventing cells from proceeding through mitosis. We previously showed that the levels of Wee-1 correlate with Vpr-mediated apoptosis. Here, we demonstrate that Vpr-induced G(2) arrest correlated with delayed degradation of Wee-1 at G(2)/M. Experimental depletion of Wee-1 by a small interfering RNA directed to wee-1 mRNA alleviated Vpr-induced G(2) arrest and allowed apparently normal progression through M into G(1). Similar results were observed when cells were arrested at G(2) following gamma irradiation. Thus, Wee-1 is integrally involved as a key cellular regulatory protein in the signal transduction pathway for HIV-1 Vpr-induced cell cycle arrest. 相似文献
995.
目的:观察新生SD大鼠原代培养皮层神经元的钙激活钾通道(Kca)在黎芦碱致神经元损伤模型上的激活、抑制效应.方法:采用细胞贴附和内面向外两种膜片钳单通道记录方法记录新生SD大鼠原代培养皮层神经元的Kca电生理活动.结果:黎芦碱在胞外可激活Kca.在有钙浴液内,细胞贴附式,钳制膜电位 30 mV,加入不同浓度黎芦碱(μmol/L:15、25、50、75),通道开放概率由0.005分别增加为0.014±0.003、0.085±0.010、0.132±0.016、0.059±0.006(P<0.01),在50μmol/L以内表现出浓度依赖性.无钙浴液内,细胞贴附式膜片上,钳制膜电位 50 mV,随药物浓度(μmol/L)增加为15、40、60、100时,通道开放概率由0.005分别增加为0.014±0.010、0.113±0.006、0.141±0.004、0 295±0.009(P<0.05).6例内面向外式膜片上,钳制膜电位 40 mV,分别加入黎芦碱25 μmol/L、50μmol/L 3 min后,通道开放概率由0.011±0.008分别增加为0.010±0.010、0.012±0.007(P>0.05).黎芦碱在胞内Kca开放概率,平均开放/关闭时间,电流幅值均无明显变化.结论:黎芦碱通过影响胞内游离钙水平间接调节Kca,在缺血缺氧早期,胞内游离钙增高激活Kca开放. 相似文献
996.
黑龙江省是肾综合征因热(HFRS)的重疫区。近年来HFRS年的发病人数曾超过万人。流行病学和血清学研究表明黑龙江省HFRS疫区主要是姬鼠型,但目前尚缺乏病毒的分子生物学资料。我们对从疫区捕获的宿主动物-黑线姬鼠肺中分离的汉坦病毒HTN261株的S基因片段的全基因序列进测定和初步分析。结果如下,HTN261株的S基因片段的全序列长为1697nt。只有一个主要的编码N蛋白的ORF,起始位置为第37nt,终止于1326nt,编码的蛋白长为429aa。没有发现存在ORF2。HTN261株的S基因片段核苷酸序列与HGTN型中的病毒株的同源性很高,而与汉坦病毒其他型的同源民生较差,从种系发生树分析来看,HNT261株归结于汉坦病毒的HTN型。在HTN型之内,HTN261株和HTN76-118株在一个分枝内,就其核苷酸和蛋白的同源性说,HT N261株和HTN76-118株的同源性分别是89%(全S基因)和98%(蛋白)。而与中国境内发现的其他汉坦病毒株Z10,HU,Chen4,NC167等基因和蛋白的同源性相对较差,汉坦病毒除具有其宿主的依赖性外,还具有其地理的簇集性。HTN261株和HTN76-118株之间S基因和N蛋白序列的变异性的差异分别为11%和2%,表明HTN261株和HTN76-118株还有不同,可能是不同的亚型。不过,尚有待于进一步研究证明。 相似文献
997.
葡萄叶绿体rbcL基因的结构分析 总被引:2,自引:0,他引:2
以玫瑰香葡萄(Vitisvinifera L.)为材料,克隆了含有叶绿体rbc L基因的3.1 kb Bam HⅠ片段,构建了该基因的限制性酶切图谱,测定了该基因的核苷酸序列。所测的核苷酸序列总长度为2004 bp,其中基因的编码区为1428 bp,编码一个含475 个氨基酸的蛋白质,其分子量约为53 kD;测定基因的5上游含启动子的部分共358 bp,包括- 10 区(TAAAAT)、- 35区(TTGCGC)和SD 序列(GGAGG);基因的3下游区共218 bp,含有3 个转录茎环终止结构。玫瑰香葡萄rbc L基因编码区的核苷酸序列与烟草、矮牵牛、菠菜、苜蓿、水稻和玉米之间的同源性分别为91.5% 、91.4% 、90.2% 、89.8% 、86.3% 和84.5% ;推导出的氨基酸序列的同源性分别为92.2% 、91.6% 、92.2% 、93.7% 、93.5% 和90.1% 。 相似文献
998.
虫花菌(Isaria farinosa(Dicks)Fr.)发酵液经超滤浓缩、乙醇沉淀、弱酸性阳离子交换树脂和弱碱性阴离子交换树脂去杂蛋白后,又经Sephadex G-150纯化得到PG。PG经醋酸纤维膜电泳、聚丙烯酰胺凝胶电泳和Sepharose 4B柱层析,证明是单一均匀的糖蛋白。PG的分子量为11.1万。PG的糖含量为92.35%、蛋白含量为7.61%。PG用气相色谱、红外光谱分析表明含有D-甘露糖、D-半乳糖,其摩尔比是5.93:1。推测PG主要含α-型糖苷键。PG对小鼠实体瘤S-180有一定的抑制作用,抑瘤率为22.2%。 相似文献
999.
1000.
Ying-Jin Yuan Jing-Chuan Li Zhi-Qiang Ge Jin-Chuan Wu 《Journal of Molecular Catalysis .B, Enzymatic》2002,18(4-6):251-260
Generation of reactive oxygen species (ROS) induced by Ce4+ in suspension cultures of Taxus cuspidata was investigated. The burst of superoxide anions (O2√−) occurred rapidly after the addition of Ce4+ and reached maximum at 4.3 h, while the total level of the cellular reactive oxygen species maintained unchanged. The intracellular superoxide dismutase (SOD) and catalase (CAT) were activated while the intra/extracellular peroxidases (PODs) were inhibited accompanying the O2√− burst. The pretreatment of the suspension cultures with diphenylene iodonium (DPI), a suicide inhibitor of the NADPH oxidase, blocked the O2√− burst, inhibiting the cell apoptosis and taxol production induced by Ce4+. These results show that NADPH oxidase played a key role in O2√− burst and O2√− served as a mediator of Ce4+ for cell apoptosis and taxol production. The pretreatments of the suspension cultures with anthracene-9-carboxylate, an ion-channel blocker, nifedipine, a Ca2+-channel blocker, neomycin, a phospholipase C (PLC) inhibitor, or suramin, a G-protein inhibitor, decreased O2√− burst induced by Ce4+. It is thus inferred that Ce4+-induced O2√− burst, which mediated cell apoptosis and taxol production by activating the ion-channels, PLC, G-proteins and NADPH oxidase. 相似文献