低频电磁场对酿酒酵母蛋白质表达及酶活性的影响
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摘要
【背景】低频电磁场(Lowfrequencyelectromagneticfield,LF-EMF)可导致一系列健康效应的事实已被普遍接受。【目的】进一步探究LF-EMF辐射对细胞蛋白质表达及相关酶活性的影响。【方法】以酿酒酵母为研究对象,采用50HzLF-EMF对其进行持续暴露辐射,随后对其胞内蛋白质表达、相关酶活性进行分析。【结果】LF-EMF辐射对酿酒酵母超氧化物歧化酶(Superoxide dismutase,SOD)、过氧化氢酶(Catalase,CAT)和苹果酸脱氢酶(Malate dehydrogenase,MDH)活性的影响存在先促进后抑制的趋势。辐射处理使SOD活性在72 h提高了43.18%,CAT活性在16 h提高了12.22%,MDH活性在20h提高了12.90%(P<0.05);而LF-EMF辐射作用对乙醇脱氢酶(Alcohol dehydrogenase,ADH)和乳酸脱氢酶(Lactatedehydrogenase,LDH)的活性并未观察到显著的影响(P>0.05)。此外,蛋白质电泳和蛋白质组学分析的结果表明,LF-EMF可改变酿酒酵母部分胞内蛋白质的表达水平,其中3种蛋白质的表达发生显著上调,9种蛋白质的表达发生显著下调(P<0.05)。【结论】LF-EMF辐射可影响酿酒酵母的部分酶活性,改变其胞内部分蛋白质的表达水平。
[Background] It is now well accepted that low frequency electromagnetic field(LF-EMF)would produce a variety of health effects. [Objective] The objective of this work is to further investigate the effects of LF-EMF radiation on cells' protein expression and enzymes activities. [Methods]Saccharomyces cerevisiae was used as the research object and was continuously exposed to 50 Hz low frequency electromagnetic field in this work, then the expression of its intracellular proteins and relevant enzyme activities were analyzed. [Results] The effect of LF-EMF treatment on the activities of SOD(superoxide dismutase), CAT(catalase) and MDH(malate dehydrogenase) in S. cerevisiae appeared a trend of firstly promoted and then inhibited. LF-EMF treatment can increase 43.18% of the activity of SOD at 72 h, 12.22% of the CAT at 16 h, and 12.90% of the MDH at 20 h(P<0.05); however, no significant effects of LF-EMF treatment on the activity of ADH(alcohol dehydrogenase) and LDH(lactate dehydrogenase) were observed(P>0.05). In addition, the results of protein electrophoresis and proteomics analysis indicated that LF-EMF could change the expression level of intracellular proteins in S. cerevisiae,including significant up-regulation of 3 proteins and significant down-regulation of 9 proteins(P<0.05).[Conclusion] LF-EMF radiation can affect the activity of relevant enzymes in S. cerevisiae, and change the expression level of some intracellular proteins.
[Background] It is now well accepted that low frequency electromagnetic field(LF-EMF)would produce a variety of health effects. [Objective] The objective of this work is to further investigate the effects of LF-EMF radiation on cells' protein expression and enzymes activities. [Methods]Saccharomyces cerevisiae was used as the research object and was continuously exposed to 50 Hz low frequency electromagnetic field in this work, then the expression of its intracellular proteins and relevant enzyme activities were analyzed. [Results] The effect of LF-EMF treatment on the activities of SOD(superoxide dismutase), CAT(catalase) and MDH(malate dehydrogenase) in S. cerevisiae appeared a trend of firstly promoted and then inhibited. LF-EMF treatment can increase 43.18% of the activity of SOD at 72 h, 12.22% of the CAT at 16 h, and 12.90% of the MDH at 20 h(P<0.05); however, no significant effects of LF-EMF treatment on the activity of ADH(alcohol dehydrogenase) and LDH(lactate dehydrogenase) were observed(P>0.05). In addition, the results of protein electrophoresis and proteomics analysis indicated that LF-EMF could change the expression level of intracellular proteins in S. cerevisiae,including significant up-regulation of 3 proteins and significant down-regulation of 9 proteins(P<0.05).[Conclusion] LF-EMF radiation can affect the activity of relevant enzymes in S. cerevisiae, and change the expression level of some intracellular proteins.
引文
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[21]Morelli A,Ravera S,Panfoli I,et al.Effects of extremely low frequency electromagnetic fields on membrane-associated enzymes[J].Archives of Biochemistry and Biophysics,2005,441(2):191-198
[22]Qiao GH,Li T,Wang KR.Molecular cloning and analysis of ubiquitin and ribosomal protein L40 fusion protein from Cryphonectria parasitica[J].Jiangsu Journal of Agricultural Sciences,2009,25(4):758-762(in Chinese)乔广行,李涛,王克荣.栗疫病菌泛素核糖体L40融合蛋白基因的克隆与分析[J].江苏农业学报,2009,25(4):758-762
[23]Erni B,Siebold C,Christen S,et al.Small substrate,big surprise:fold,function and phylogeny of dihydroxyacetone kinases[J].Cellular and Molecular Life Sciences CMLS,2006,63(7/8):890-900
[24]Ju SL,Shaltiel G,Shamir A,et al.Human1-D-myo-inositol-3-phosphate synthase is functional in yeast[J].Journal of Biological Chemistry,2004,279(21):21759-21765
[25]Zhong Q,Greenberg ML.Regulation of phosphatidylglycerophosphate synthase by inositol in Saccharomyces cerevisiae is not at the level of PGS1 m RNAabundance[J].Journal of Biological Chemistry,2003,278(36):33978-33984
[26]Cai XH,Ren HF,Meng YS,et al.Effects of low frequency electrostatic field radiation on growth and metabolism of Saccharomyces cerevisiae[J].Science and Technology of Food Industry,2018,39(2):124-127,133(in Chinese)蔡杏华,任浩锋,蒙永双,等.低频静电场辐射对酿酒酵母生长代谢的影响[J].食品工业科技,2018,39(2):124-127,133
[2]Wertheimer N,Leeper E.Electrical wiring configurations and childhood cancer[J].American Journal of Epidemiology,1979,109(3):273-284
[3]IARC Working Group on the Evaluation of Carcinogenic Risks to Humans.Non-Ionizing Radiation,Part 1:Static and Extremely Low-Frequency(ELF)Electric and Magnetic Fields[M].Lyon,France:IARC,2002:285
[4]Zhang PP,Yin RC,Wu LF,et al.Non-thermal bioeffects of static and extremely low frequency electromagnetic fields[J].Journal of Biomedical Engineering,2007,24(6):1411-1415(in Chinese)张萍萍,尹若春,吴丽芳,等.静态和极低频电磁场非热生物效应的研究动态[J].生物医学工程学杂志,2007,24(6):1411-1415
[5]Liu Y,Weng EQ,Dai YQ,et al.Biological effects of extremely low frequency electromagnetic fields on soybean seeds during germination[J].Chinese Journal of Applied&Environmental Biology,2002,8(5):482-484(in Chinese)刘赟,翁恩琪,戴雅奇,等.极低频电磁场对萌发期大豆的生物学效应[J].应用与环境生物学报,2002,8(5):482-484
[6]Amara S,Douki T,Garrel C,et al.Effects of static magnetic field and cadmium on oxidative stress and DNA damage in rat cortex brain and hippocampus[J].Toxicology and Industrial Health,2011,27(2):99-106
[7]Panagopoulos DJ,Karabarbounis A,Lioliousis C.ELF alternating magnetic field decreases reproduction by DNA damage induction[J].Cell Biochemistry and Biophysics,2013,67(2):703-716
[8]Cappelli S,Penco A,Mannini B,et al.Effect of molecular chaperones on aberrant protein oligomers in vitro:super-versus sub-stoichiometric chaperone concentrations[J].Biological Chemistry,2016,397(5):401-415
[9]Patruno A,Tabrez S,Pesce M,et al.Effects of extremely low frequency electromagnetic field(ELF-EMF)on catalase,cytochrome P450 and nitric oxide synthase in erythro-leukemic cells[J].Life Sciences,2015,121:117-123
[10]Song J,Lin JQ,Jin Y,et al.A new population growth model based on the time dependent changes of the specific growth rate[J].Microbiology China,2007,34(5):836-838(in Chinese)宋健,林建群,金燕,等.以比生长速率时间曲线为基础的生物群体生长数学模型[J].微生物学通报,2007,34(5):836-838
[11]Carlsson N,Borde A,W?lfel S,et al.Quantification of protein concentration by the Bradford method in the presence of pharmaceutical polymers[J].Analytical Biochemistry,2011,411(1):116-121
[12]Fibla J,Gonzàlez-Duarte R.Colorimetric assay to determine alcohol dehydrogenase activity[J].Journal of Biochemical and Biophysical Methods,1993,26(1):87-93
[13]Zhang HF.Discussion on definition of activity unit of enzyme[J].Chinese Science and Technology Terms Journal,2005,7(1):38(in Chinese)张华峰.关于酶活力单位定义[J].科技术语研究,2005,7(1):38
[14]Petschnigg J,Groisman B,Kotlyar M,et al.The mammalianmembrane two-hybrid assay(MAMTH)for probing membrane-protein interactions in human cells[J].Nature Methods,2014,11(5):585-592
[15]Xiong Y,Tang XF,Meng QS,et al.Differential expression analysis of the broiler tracheal proteins responsible for the immune response and muscle contraction induced by high concentration of ammonia using iTRAQ-coupled 2DLC-MS/MS[J].Science China Life Sciences,2016,59(11):1166-1176(in Chinese)熊嫣,唐湘方,孟庆石,等.高氨刺激下参与免疫应答和肌肉收缩过程的肉鸡气管蛋白差异表达--基于iTRAQ标记技术的差异蛋白质组学研究[J].中国科学:生命科学,2016,46(11):1267-1276
[16]Boutet E,Lieberherr D,Tognolli M,et al.UniProtKB/Swiss-Prot,the manually annotated section of the UniProt knowledgebase:how to use the entry view[A]//Edwards D.Plant Bioinformatics:Methods and Protocols[M].New York,NY:Humana Press,2016:23-54
[17]Wang BH.Simulation comparison of two fisher exact test algorithms[J].Chinese Journal of Health Statistics,2005,22(2):114-115(in Chinese)王斌会.两种Fisher精确检验算法的模拟比较[J].中国卫生统计,2005,22(2):114-115
[18]Sadeghipour R,Ahmadian S,Bolouri B,et al.Effects of extremely low-frequency pulsed electromagnetic fields on morphological and biochemical properties of human breast carcinoma cells(T47D)[J].Electromagnetic Biology and Medicine,2012,31(4):425-435
[19]Chen GD,Xu ZP.Global protein expression in response to extremely low frequency magnetic fields[A]//Leszczynski D.Radiation Proteomics:the Effects of Ionizing and Non-Ionizing Radiation on Cells and Tissues[M].Dordrecht:Springer,2013:107-110
[20]Ciejka E,Kleniewska P,Skibska B,et al.Effects of extremely low frequency magnetic field on oxidative balance in brain of rats[J].Journal of Physiology and Pharmacology:An Official Journal of the Polish Physiological Society,2011,62(6):657-661
[21]Morelli A,Ravera S,Panfoli I,et al.Effects of extremely low frequency electromagnetic fields on membrane-associated enzymes[J].Archives of Biochemistry and Biophysics,2005,441(2):191-198
[22]Qiao GH,Li T,Wang KR.Molecular cloning and analysis of ubiquitin and ribosomal protein L40 fusion protein from Cryphonectria parasitica[J].Jiangsu Journal of Agricultural Sciences,2009,25(4):758-762(in Chinese)乔广行,李涛,王克荣.栗疫病菌泛素核糖体L40融合蛋白基因的克隆与分析[J].江苏农业学报,2009,25(4):758-762
[23]Erni B,Siebold C,Christen S,et al.Small substrate,big surprise:fold,function and phylogeny of dihydroxyacetone kinases[J].Cellular and Molecular Life Sciences CMLS,2006,63(7/8):890-900
[24]Ju SL,Shaltiel G,Shamir A,et al.Human1-D-myo-inositol-3-phosphate synthase is functional in yeast[J].Journal of Biological Chemistry,2004,279(21):21759-21765
[25]Zhong Q,Greenberg ML.Regulation of phosphatidylglycerophosphate synthase by inositol in Saccharomyces cerevisiae is not at the level of PGS1 m RNAabundance[J].Journal of Biological Chemistry,2003,278(36):33978-33984
[26]Cai XH,Ren HF,Meng YS,et al.Effects of low frequency electrostatic field radiation on growth and metabolism of Saccharomyces cerevisiae[J].Science and Technology of Food Industry,2018,39(2):124-127,133(in Chinese)蔡杏华,任浩锋,蒙永双,等.低频静电场辐射对酿酒酵母生长代谢的影响[J].食品工业科技,2018,39(2):124-127,133