大麻CBDA1基因的生物信息学分析
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摘要
[目的]研究大麻二酚酸合成酶基因(CBDA1)编码的蛋白质序列所包含的生物学信息。[方法]利用生物信息学在线工具及软件分析大麻二酚酸合成酶(CBDAS)的理化性质、亲/疏水性、跨膜结构、信号肽、motif结构域及空间结构等。[结果]CBDAS由544个氨基酸组成,分子量为62 168.42,理论等电点为8.81,是一种稳定的亲水性分泌蛋白,N末端包含1个由28个氨基酸残基组成的信号肽,该蛋白的亚细胞定位在胞外。CBDAS属于氧化还原酶家族,FDA是该酶活性的必需辅因子,CBDAS蛋白中只含有1个低复杂度区域,含有23个磷酸化位点和6个N-糖基化位点,其二级结构主要由α-螺旋、β-转角和无规卷曲组成,三级结构与四氢大麻酚酸合成酶(THCAS)同源性最高。[结论]该研究结果可为今后深入研究CBDAS蛋白的结构特征和功能提供理论参考。
[Objective]To study the biological information contained in the protein sequence encoded by cannabidiolic acid synthase gene( CBDA1). [Method]The physicochemical properties,hydrophilicity and hydrophobicity,transmembrane structure domain,the signal peptide,the motif and the spatial structure of cannabidiolic acid synthase( CBDAS) were analyzed by bioinformatic sever and online tools. [Result]CBDAS consisted of 544 amino acids with molecular weight of 62 168. 42 and a theoretical isoelectric point of 8. 81. It was a stable hydrophilic secretory protein and the N-terminal contained a signal peptide of 28 amino acid residues,moreover,the subcellular localization of CBDAS was extracellular. It was presumed that CBDAS belonged to the family of oxidoreductases and FDA was a necessary cofactor for the activity of CBDAS. In addtion,CBDAS contained one low complexity region,twenty three phosphorylation sites and six N-glycosylation sites. The secondary structure mainly included α-helix,β-turn and random coil. Furthermore,the tertiary structure was the most homologous to tetrahydrocannabinol synthase( THCAS). [Conclution]The results could provide a theoretical reference for further study of the structure and functions of CBDAS.
[Objective]To study the biological information contained in the protein sequence encoded by cannabidiolic acid synthase gene( CBDA1). [Method]The physicochemical properties,hydrophilicity and hydrophobicity,transmembrane structure domain,the signal peptide,the motif and the spatial structure of cannabidiolic acid synthase( CBDAS) were analyzed by bioinformatic sever and online tools. [Result]CBDAS consisted of 544 amino acids with molecular weight of 62 168. 42 and a theoretical isoelectric point of 8. 81. It was a stable hydrophilic secretory protein and the N-terminal contained a signal peptide of 28 amino acid residues,moreover,the subcellular localization of CBDAS was extracellular. It was presumed that CBDAS belonged to the family of oxidoreductases and FDA was a necessary cofactor for the activity of CBDAS. In addtion,CBDAS contained one low complexity region,twenty three phosphorylation sites and six N-glycosylation sites. The secondary structure mainly included α-helix,β-turn and random coil. Furthermore,the tertiary structure was the most homologous to tetrahydrocannabinol synthase( THCAS). [Conclution]The results could provide a theoretical reference for further study of the structure and functions of CBDAS.
引文
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[25]叶强,金晓琴,刘伟娜,等.植物蛋白质N-糖基化修饰研究进展[J].浙江师范大学学报(自然科学版),2016,39(1):80-86.
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[27]SEN T Z,JERNIGAN R L,GARNIER J,et al.GOR V server for protein secondary structure prediction[J].Bioinformatics,2005,21(11):2787-2788.
[28]SHOYAMA Y,TAMADA T,KURIHARA K,et al.Structure and function ofΔ1-tetrahydrocannabinolic acid(THCA)synthase,the enzyme controlling the psychoactivity of Cannabis sativa[J].Journal of molecular biology,2012,423(1):96-105.
[2]GAONI Y,MECHOULAM R.Isolation,structure,and partial synthesis of an active constituent of hashish[J].Journal of the American chemical society,1964,86(8):1646-1647.
[3]ABRAMS D I,GUZMAN M.Cannabis in cancer care[J].Clinical pharmacology and therapeutics,2015,97(6):575-586.
[4]TAURA F,MORIMOTO S,SHOYAMA Y,et al.First direct evidence for the mechanism ofΔ1-tetrahydrocannabinolic acid biosynthesis[J].Journal of the American chemical society,1995,117(38):9766-9767.
[5]SIRIKANTARAMAS S,MORIMOTO S,SHOYAMA Y,et al.The gene controlling marijuana psychoactivity[J].Journal of biological chemistry,2004,279(38):39767-39774.
[6]SHOYAMA Y,TAMADA T,KURIHARA K,et al.Structure and function ofΔ1-tetrahydrocannabinolic acid(THCA)synthase,the enzyme controlling the psychoactivity of Cannabis sativa[J].Journal of molecular biology,2012,423(1):96-105.
[7]KLEIN B D,JACOBSON C A,METCALF C S,et al.Evaluation of cannabidiol in animal seizure models by the Epilepsy Therapy Screening Program(ETSP)[J].Neurochemical research,2017,42(7):1939-1948.
[8]BORRELLI F,AVIELLO G,ROMANO B,et al.Cannabidiol,a safe and non-psychotropic ingredient of the marijuana plant Cannabis sativa,is protective in a murine model of colitis[J].Journal of molecular medicine,2009,87:1111-1121.
[9]SYED Y Y,MCKEAGE K,SCOTT L J.Delta-9-tetrahydrocannabinol/cannabidiol(Sativex):A review of its use in patients with moderate to severe spasticity due to multiple sclerosis[J].Botany and biotechnology,2014,74(5):563-578.
[10]BLESSING E M,STEENKAMP M M,MANZANARES J,et al.Cannabidiol as a potential treatment for anxiety disorders[J].Neurotherapeutics,2015,12(4):825-836.
[11]TAURA F,MORIMOTO S,SHOYAMA Y,et al.Purification and characterization of cannabidiolic-acid from Cannabis sativa L.[J].Journal of biological chemistry,1996,271(29):17411-17416.
[12]TAURA F,SIRIKANTARAMAS S,SHOYAMA Y,et al.Cannabidiolicacid synthase,the chemotype-determining enzyme in the fiber-type Cannabis sativa[J].FEBS letters,2007,581(16):2929-2934.
[13]于欣,杨震,楚元奎,等.IL-6基因结构和功能生物信息学预测[J].国际检验医学杂志,2016,37(21):2959-2960,2963.
[14]丁帅,熊勇,李正涛,等.菊花rbc L基因电子克隆及生物信息学、适应性进化分析[J].种子,2015,34(10):24-30.
[15]KROGH A,LARSSON B,VON HEIJNE G,et al.Predicting transmembrane protein topology with a hidden Markov model:Application to complete genomes[J].Journal of molecular biology,2001,305(3):567-580.
[16]YAN S,WU G.Signal peptide of cellulase[J].Applied microbiology and biotechnology,2014,98(12):5329-5362.
[17]IZARD J W,DOUGHTY M B,KENDALL D A.Physical and conformational properties of synthetic idealized signal sequences parallel their biological functional[J].Biochemistry,1995,34(31):9904-9912.
[18]PETERSEN T N,BRUNAK S,VON HEIJNE G,et al.Signal P 4.0:Discriminating signal peptides from transmembrane regions[J].Nature methods,2011,8(10):785-786.
[19]张松,黄波,夏学峰,等.蛋白质亚细胞定位的生物信息学研究[J].生物化学与生物物理进展,2007,34(6):573-579.
[20]CUNANE L M,CHEN Z W,SHAMALA N,et al.Structures of the flavocytochrome p-cresol methylhydroxylase and its enzyme-substrate complex:Gated substrate entry and proton relays support the proposed catalytic mechanism[J].Journal of molecular biology,2000,295(2):357-374.
[21]FRAAIJE M W,MATTEVI A.Flavoenzymes:Diverse catalysts with recurrent features[J].Trends in biochemical sciences,2000,25(3):126-132.
[22]MCINTIRE W,EDMONDSON D E,HOPPER D J,et al.8 alpha-(O-Tyrosyl)flavin adenine dinucleotide,the prosthetic group of bacterial p-cresol methylhydroxylase[J].Biochemistry,1981,20(11):3068-3075.
[23]EISENHABER B,EISENHABER F.Prediction of posttranslational modification of proteins from their amino acid sequence[J].Methods in molecular biology,2010,609:365-384.
[24]刘秋林,钟月仙,万伟峰,等.植物磷酸化蛋白质组学研究进展[J].福建农林大学学报(自然科学版),2015,44(3):225-231.
[25]叶强,金晓琴,刘伟娜,等.植物蛋白质N-糖基化修饰研究进展[J].浙江师范大学学报(自然科学版),2016,39(1):80-86.
[26]LEROUGE P,CABANES-MACHETEAU M,RAYON C,et al.N-Glycoprotein biosynthesis in plants:Recent developments and future trends[J].Plant molecular biology,1998,38(1/2):31-48.
[27]SEN T Z,JERNIGAN R L,GARNIER J,et al.GOR V server for protein secondary structure prediction[J].Bioinformatics,2005,21(11):2787-2788.
[28]SHOYAMA Y,TAMADA T,KURIHARA K,et al.Structure and function ofΔ1-tetrahydrocannabinolic acid(THCA)synthase,the enzyme controlling the psychoactivity of Cannabis sativa[J].Journal of molecular biology,2012,423(1):96-105.