杨铁,龙高群,张春林
YANG Tie,LONG Gaoqun,ZHANG Chunlin

• 1:贵阳医学院生物学教研室

摘要(Abstract)：

目的:通过生物信息学方法预测德国小蠊致敏原蛋白Bla g 8的三维结构和抗原表位,为蟑螂引起人类变态反应性疾病的诊断和治疗提供线索。方法:在美国国立生物信息中心(NCBI)上获取Bla g 8致敏原蛋白序列,通过序列比对(BLAST)得到相似序列,应用MEGA6.0构建同源进化树;联合应用DNAStar多种方法预测其抗原表位,使用Modeller 9.12同源建模后经Gromacs能量优化获取稳定的致敏原蛋白。结果:德国小蠊Bla g 8与美洲大蠊肌球蛋白轻链在进化上具有较近的亲缘关系;Bla g 8为含有2个EF手钙结合蛋白位点的α结构疏水性蛋白,主要抗原表位集中于1~24,27~50,55~65,69~91,92~110,121~132,138~153,159~180,191~208区域;预测并经能量优化后得到稳定的三维结构。结论:通过预测Bla g 8结构及抗原表位,为筛选低致敏原衍生物提供了分子基础。
Objective: To predict the structures and antigenic epitopes of allergen Bla g 8 of German cockroach with bioinfomatics methods,and so as to provide clues for diagnosis and treatment of human allergic diseases caused by cockroach. Methods: The sequences of Bla g 8 were obtained from USA national center for biotechnology information( NCBI). Sequence alignment( BLAST) was applied to search for the similar protein of Bla g 8. MEGA6. 0 was used for constructing phylogenetic tree. Antigenic epitopes of Bla g 8 were predicted by the software of DNAStar. Homologous modeling was carried out with the Modeler 9. 12,and stable allergen was obtained through energy optimization with Gromacs. Results: Bla g 8 and myosin light chain of Periplaneta americana had a close genetic relationship in evolution. Bla g 8 had hydrophobic proteins of alpha secondary structure containing 2 EF hand calcium binding sites. Its antigenic epitope mostly located on 1 ~ 24,27 ~ 50,55 ~ 65,69 ~ 91,92 ~110,121 ~ 132,138 ~ 153,159 ~ 180 and 191 ~ 208 regions. Stable three dimensional structure was obtained through energy optimization. Conclusions: It provides molecular basis for screening hypoallergenic variant of cockroach allergens through predicting the structures and antigenic epitopes of Bla g 8.

关键词(KeyWords)： 蠊;致敏原;Bla g 8;抗原;表位;蛋白质序列分析
Blattella germanica;sensitinogen;Bla g 8;antigen;epitope;protein sequence analysis

Abstract:

Keywords:

基金项目(Foundation): 贵州省攻关项目[黔科合S系(20071038)];; 贵州省教育厅自然科学研究项目[黔教合KY字(2012)039];; 贵阳医学院研究生创新计划专项基金

作者(Author): 杨铁,龙高群,张春林
YANG Tie,LONG Gaoqun,ZHANG Chunlin

DOI: 10.19367/j.cnki.1000-2707.2014.05.016

参考文献(References)：

• [1]Liu Z,Bai Y,Ji K,et al.Detection of Dermatophagoides farinae in the dust of air conditioning filters[J].Int Arch Allergy Immunol,2007(1):85-90.
• [2]D'Amato G,Baena-Cagnani CE,Cecchi L,et al.Climate change,air pollution and extreme events leading to increasing prevalence of allergic respiratory diseases[J].Multidiscip Respir Med,2013(1):12.
• [3]Pawankar R,Canonica GW,Holgate ST,et al.WAO white book on allergy[J].Milwaukee,WI:World Allergy Organization,2011(3):156-157.
• [4]Bernton HS,Brown H.Insect Allergy-Preliminary Studies of the Cockroach[J].J Allergy,1964(35):506-513.
• [5]Van der Kamp MW,Mulholland AJ.Combined quantum mechanics/molecular mechanics(QM/MM)methods in computational enzymology[J].Biochemistry,2013(16):2708-2728.
• [6]Barbosa MC,Santos AB,Ferriani VP,et al.Efficacy of recombinant allergens for diagnosis of cockroach allergy in patients with asthma and/or rhinitis[J].Int Arch Allergy Immunol,2013(3):213-219.
• [7]Arruda LK,Pomes A.Every cockroach is beautiful to its mother[J].Int Arch Allergy Immunol,2013(4):289-292.
• [8]Gore JC,Schal C.Cockroach allergen biology and mitigation in the indoor environment[J].Annu Rev Entomol,2007(52):439-463.
• [9]Sookrung N,Chaicumpa W.A revisit to cockroach allergens[J].Asian Pac J Allergy Immunol,2010(2-3):95-106.
• [10]Meiler F,Zumkehr J,Klunker S,et al.In vivo switch to IL-10-secreting T regulatory cells in high dose allergen exposure[J].J Exp Med,2008(12):2887-2898.
• [11]Gardner LM,O'Hehir RE,Rolland JM.High dose allergen stimulation of T cells from house dust mite-allergic subjects induces expansion of IFN-gamma+T Cells,apoptosis of CD4+IL-4+T cells and T cell anergy[J].Int Arch Allergy Immunol,2004(1):1-13.
• [12]Patterson ML,Slater JE.Characterization and comparison of commercially available German and American cockroach allergen extracts[J].Clin Exp Allergy,2002(5):721-727.
• [13]Moverare R,Elfman L,Vesterinen E,et al.Development of new IgE specificities to allergenic components in birch pollen extract during specific immunotherapy studied with immunoblotting and Pharmacia CAP System[J].Allergy,2002(5):423-430.
• [14]龙高群,张春林.德国小蠊变应原Bla g 8的克隆表达及进化分析[J].生物技术通报,2012(9):173-178.
• [15]Gadermaier G,Jahn-Schmid B,Vogel L,et al.Targeting the cysteine-stabilized fold of Art v 1 for immunotherapy of Artemisia pollen allergy[J].Mol Immunol,2010(6):1292-1298.
• [16]Swoboda I,Bugajska-Schretter A,Linhart B,et al.A recombinant hypoallergenic parvalbumin mutant for immunotherapy of IgE-mediated fish allergy[J].J Immunol,2007(10):6290-6296.
• [17]King N,Helm R,Stanley JS,et al.Allergenic characteristics of a modified peanut allergen[J].Mol Nutr Food Res,2005(10):963-971.
• [18]Kapcha LH,Rossky PJ.A simple atomic-level hydrophobicity scale reveals protein interfacial structure[J].J Mol Biol,2014(2):484-498.
• [19]Huang JT,Huang WS,Huang R,et al.How the folding rates of two-and multistate proteins depend on the amino acid properties[J].Proteins,2014(6):1-8.
• [20]Jameson BA,Wolf H.The antigenic index:a novel algorithm for predicting antigenic determinants[J].Comput Appl Biosci,1988(1):181-186.
• [21]肖杰,吴序栎,王琳琳,等.花生profilin蛋白的生物信息学分析[J].免疫学杂志,2011(2):158-161.
• [22]Knol EF.Requirements for effective IgE cross-linking on mast cells and basophils[J].Mol Nutr Food Res,2006(7):620-624.

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