海州香薷不同种群细胞壁转化酶蛋白结构与分子模拟对接
|
摘要
为研究铜胁迫下海州香薷(Elsholtzia haichowensis Sun)非矿区种群细胞壁转化酶(Eh Ncw INV)和矿区种群细胞壁转化酶(Eh Ccw INV)活性差异的机理,运用在线分析工具对细胞壁转化酶基因推导的氨基酸序列进行分析,利用SWISS-MODEL进行同源建模,并将细胞壁转化酶与蔗糖分子进行模拟对接。结果表明,海州香薷两个种群细胞壁转化酶蛋白质在443位点和538位点处存在氨基酸趋异位点,模拟的3D结构非常相似,仅在趋异位点Leu433/Pro433和Tyr538/His538处有差别。Eh Ncw INV、Eh Ccw INV和模拟突变体(Eh Ncw INV-L433P和Eh Ncw INV-Y538H)与蔗糖分子对接形成的催化活性中心构象基本一致,但在空间位置上存在细微差异,氨基酸趋异可能是造成两个种群细胞壁转化酶活性差异的原因之一。
It was to study the mechanism of enzyme activity difference of cell wall invertase from different populations,one from non-mine population(Eh Ncw INV),the other from mine population(Eh Ccw INV) in Elsholtzia haichowensis Sun under copper stress. The amino acid sequences of cell wall invertase were analyzed by Ex PASy(http://au.expasy.org/tools/). The threedimensional structure was constructed by homologous modeling. The structures of cell wall invertase from two populations of E.haichowensis Sun and non-mine population with each single point mutation in complex with sucrose were simulated. The results showed that there were two divergent amino acids at position 443 and 538 between Eh Ncw INV and Eh Ccw INV. The threedimensional structures were exactly similar between Eh Ncw INV and Eh Ccw INV. It showed difference at Leu433/Pro433 and Tyr538/His538,which were divergent sites. The structures of catalytic active center of Eh Ncw INV,Eh Ccw INV,and simulated mutants(Eh Ncw INV-L433 P,Eh Ncw INV-Y538H) binding with sucrose showed no significant differences. However,there were differences on spatial position. It might be one of the reasons for the difference of cell wall invertase activity between two populations.
It was to study the mechanism of enzyme activity difference of cell wall invertase from different populations,one from non-mine population(Eh Ncw INV),the other from mine population(Eh Ccw INV) in Elsholtzia haichowensis Sun under copper stress. The amino acid sequences of cell wall invertase were analyzed by Ex PASy(http://au.expasy.org/tools/). The threedimensional structure was constructed by homologous modeling. The structures of cell wall invertase from two populations of E.haichowensis Sun and non-mine population with each single point mutation in complex with sucrose were simulated. The results showed that there were two divergent amino acids at position 443 and 538 between Eh Ncw INV and Eh Ccw INV. The threedimensional structures were exactly similar between Eh Ncw INV and Eh Ccw INV. It showed difference at Leu433/Pro433 and Tyr538/His538,which were divergent sites. The structures of catalytic active center of Eh Ncw INV,Eh Ccw INV,and simulated mutants(Eh Ncw INV-L433 P,Eh Ncw INV-Y538H) binding with sucrose showed no significant differences. However,there were differences on spatial position. It might be one of the reasons for the difference of cell wall invertase activity between two populations.
引文
[1]ROLLAND F,BAENA-GONZALEZ E,SHEEN J.Sugar sensing and signaling in plants:Conserved and novel mechanisms[J].Annual Review of Plant Biology,2006,58:675-709.
[2]LI Z,PALMER W M,MARTIN A P,et al.High invertase activity in tomato reproductive organs correlates with enhanced sucrose import into,and heat tolerance of young fruit[J].Journal of Experimental Botany,2011,63(3):1155-1166.
[3]STURM A.Invertases:Primary structures,functions,and roles in plant development and sucrose partitioning[J].Plant Physiology,1999,121(1):1-8.
[4]RUAN Y L,JIN Y,YANG Y J,et al.Sugar input,metabolism,and signaling mediated by invertase:Roles in development,yield potential,and response to drought and heat[J].Molecular Plant,2010,3(6):942-955.
[5]柯文山,席红安,杨毅,等.大冶铜绿山矿区海州香薷(Elsholtzia haichowensis)植物地球化学特征分析[J].生态学报,2001,21(6):907-912.
[6]谢学锦,徐邦梁.铜矿指示植物——海州香薷[J].地质学报,1952,32(4):360-368.
[7]LIU J,XIONG Z T.Differences in accumulation and physiological response to copper stress in three populations of Elsholtzia haichowensis S.[J].Water Air and Soil Pollution,2005,168(1):5-16.
[8]熊治廷.植物抗污染进化及其遗传生态学代价[J].生态学杂志,1997,16(1):53-57.
[9]YANG M J,YANG X E,R魻MHELD V.Growth and nutrient composition of Elsholtzia splendens Nakai under copper toxicity[J].Journal of Plant Nutrition,2002,25(7):1359-1375.
[10]CAI S W,XIONG Z T,LI L,et al.Differential responses of root growth,acid invertase activity and transcript level to copper stress in two contrasting populations of Elsholtzia haichowensis[J].Ecotoxicology,2014,23(1):76-91.
[11]VERHAEST M,LAMMENS W,LE ROY K,et al.X-ray diffraction structure of a cell-wall invertase from Arabidopsis thaliana[J].Acta Crystallographica Section D Biological Crystallography,2006,62(12):1555-1563.
[12]LAMMENS W,LE ROY K,VAN LAERE A,et al.Crystal structures of Arabidopsis thaliana cell-wall invertase mutants in complex with sucrose[J].Journal of Molecular Biology,2008,377(2):378-385.
[13]DAVIES G,HENRISSAT B.Structures and mechanisms of glycosyl hydrolases[J].Structure,1995,3(9):853-859.
[14]HUANG W X,CAO Y,HUANG L J,et al.Differential expression of acid invertase genes in roots of metallicolous and non-metallicolous populations of Rumex japonicus under copper stress[J].Chemosphere,2011,84(10):1432-1439.
[15]DRAIE R,P魪RON T,POUVREAU J,et al.Invertases involved in the development of the parasitic plant Phelipanche ramosa:Characterization of the dominant soluble acid isoform,Pr SAI1[J].Molecular Plant Pathology,2011,12(7):638-652.
[16]TYMOWSKA-LALANNE Z,KREIS M.The plant invertases:Physiology,biochemistry and molecular biology[J].Advances in Botanical Research,1998,28:71-117.
[17]RITSEMA T,SMEEKENS S C M.Engineering fructan metabolism in plants[J].Journal of Plant Physiology,2003,160(7):811-820.
[18]FRIDMAN E,CARRARI F,LIU Y,et al.Zooming in on a quantitative trait for tomato yield using interspecific introgressions[J].Science,2004,305(5691):1786-1789.
[19]GOETZ M,ROITSCH T.Identification of amino acids essential for enzymatic activity of plant invertases[J].Journal of Plant Physiology,2000,157(5):581-585.
[20]LE ROY K,LAMMENS W,VERHAEST M,et al.Unraveling the difference between invertases and fructan exohydrolases:A single mmino acid(Asp-239)substitution transforms Arabidopsis cell wall invertase1 into a fructan 1-exohydrolase[J].Plant Physiology,2007,145(3):616-625.
[21]M魣TRAI J,LAMMENS W,JONCKHEER A,et al.An alternate sucrose binding mode in the E203Q Arabidopsis invertase mutant:An X-ray crystallography and docking study[J].Proteins:Structure,Function,and Bioinformatics,2008,71(2):552-564.
[22]CARLSON S J,SHANKER S,CHOUREY P S.A point mutation at the Miniature 1 seed locus reduces levels of the encoded protein,but not its m RNA,in maize[J].Molecular and General Genetics,2000,263(2):367-373.
[2]LI Z,PALMER W M,MARTIN A P,et al.High invertase activity in tomato reproductive organs correlates with enhanced sucrose import into,and heat tolerance of young fruit[J].Journal of Experimental Botany,2011,63(3):1155-1166.
[3]STURM A.Invertases:Primary structures,functions,and roles in plant development and sucrose partitioning[J].Plant Physiology,1999,121(1):1-8.
[4]RUAN Y L,JIN Y,YANG Y J,et al.Sugar input,metabolism,and signaling mediated by invertase:Roles in development,yield potential,and response to drought and heat[J].Molecular Plant,2010,3(6):942-955.
[5]柯文山,席红安,杨毅,等.大冶铜绿山矿区海州香薷(Elsholtzia haichowensis)植物地球化学特征分析[J].生态学报,2001,21(6):907-912.
[6]谢学锦,徐邦梁.铜矿指示植物——海州香薷[J].地质学报,1952,32(4):360-368.
[7]LIU J,XIONG Z T.Differences in accumulation and physiological response to copper stress in three populations of Elsholtzia haichowensis S.[J].Water Air and Soil Pollution,2005,168(1):5-16.
[8]熊治廷.植物抗污染进化及其遗传生态学代价[J].生态学杂志,1997,16(1):53-57.
[9]YANG M J,YANG X E,R魻MHELD V.Growth and nutrient composition of Elsholtzia splendens Nakai under copper toxicity[J].Journal of Plant Nutrition,2002,25(7):1359-1375.
[10]CAI S W,XIONG Z T,LI L,et al.Differential responses of root growth,acid invertase activity and transcript level to copper stress in two contrasting populations of Elsholtzia haichowensis[J].Ecotoxicology,2014,23(1):76-91.
[11]VERHAEST M,LAMMENS W,LE ROY K,et al.X-ray diffraction structure of a cell-wall invertase from Arabidopsis thaliana[J].Acta Crystallographica Section D Biological Crystallography,2006,62(12):1555-1563.
[12]LAMMENS W,LE ROY K,VAN LAERE A,et al.Crystal structures of Arabidopsis thaliana cell-wall invertase mutants in complex with sucrose[J].Journal of Molecular Biology,2008,377(2):378-385.
[13]DAVIES G,HENRISSAT B.Structures and mechanisms of glycosyl hydrolases[J].Structure,1995,3(9):853-859.
[14]HUANG W X,CAO Y,HUANG L J,et al.Differential expression of acid invertase genes in roots of metallicolous and non-metallicolous populations of Rumex japonicus under copper stress[J].Chemosphere,2011,84(10):1432-1439.
[15]DRAIE R,P魪RON T,POUVREAU J,et al.Invertases involved in the development of the parasitic plant Phelipanche ramosa:Characterization of the dominant soluble acid isoform,Pr SAI1[J].Molecular Plant Pathology,2011,12(7):638-652.
[16]TYMOWSKA-LALANNE Z,KREIS M.The plant invertases:Physiology,biochemistry and molecular biology[J].Advances in Botanical Research,1998,28:71-117.
[17]RITSEMA T,SMEEKENS S C M.Engineering fructan metabolism in plants[J].Journal of Plant Physiology,2003,160(7):811-820.
[18]FRIDMAN E,CARRARI F,LIU Y,et al.Zooming in on a quantitative trait for tomato yield using interspecific introgressions[J].Science,2004,305(5691):1786-1789.
[19]GOETZ M,ROITSCH T.Identification of amino acids essential for enzymatic activity of plant invertases[J].Journal of Plant Physiology,2000,157(5):581-585.
[20]LE ROY K,LAMMENS W,VERHAEST M,et al.Unraveling the difference between invertases and fructan exohydrolases:A single mmino acid(Asp-239)substitution transforms Arabidopsis cell wall invertase1 into a fructan 1-exohydrolase[J].Plant Physiology,2007,145(3):616-625.
[21]M魣TRAI J,LAMMENS W,JONCKHEER A,et al.An alternate sucrose binding mode in the E203Q Arabidopsis invertase mutant:An X-ray crystallography and docking study[J].Proteins:Structure,Function,and Bioinformatics,2008,71(2):552-564.
[22]CARLSON S J,SHANKER S,CHOUREY P S.A point mutation at the Miniature 1 seed locus reduces levels of the encoded protein,but not its m RNA,in maize[J].Molecular and General Genetics,2000,263(2):367-373.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |