Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (1): 154-161.doi: 10.3864/j.issn.0578-1752.2013.01.018

• ANIMAL SCIENCE·RESOURCE INSECT • Previous Articles     Next Articles

Interpretation of Odorant Binding Function and Mode of General Odorant Binding Protein ASP2 in Chinese Honeybee (Apis cerana cerana)

 LI  Hong-Liang, ZHANG  Lin-Ya, ZHUANG  Shu-Lin, NI  Cui-Xia, HAN  Bao-Yu, SHANG  Han-Wu   

  1. College of Life Sciences, China Jiliang University/Zhejiang Provincial Key Laboratory of Biometrology and Inspection & Quarantine, Hangzhou 310018
  • Received:2012-05-02 Online:2013-01-01 Published:2012-08-16

PDF

945

Cited

14

Abstract: 【Objective】 The objective of this study is to research the binding function and mode of different odors with general odorant binding protein AcerASP2 of Chinese honeybees (Apis cerana cerana). 【Method】 With the optimization of induction conditions, the purified recombinant AcerASP2 protein was obtained, then the competitive fluorescence assay was used to determine the binding function of AcerASP2 with odors having different structures, finally the homology modeling and molecular docking were applied to elucidate the binding mode and mechanism. 【Result】After the soluble recombinant AcerASP2 protein purified, in the competitive fluorescence assay between AcerASP2 and 1-NPN, the dissociation constants K1-NPN and the number of binding sites n were 7.38 μmol•L-1 and 1.0321, respectively. In selective 7 kinds of odors, the affinity of 4-allylveratrole seemed to be the strongest, and the IC50 and dissociation constant KD were 7.09 and 3.46 μmol•L-1, respectively. The molecular docking results showed that AcerASP2 had one elongated pocket-like hydrophobic cavity, 4-allylveratrole just existed in the cavity, and two hydrogen bonds were found with Lys74 of AcerASP2. 【Conclusion】Because of the particular spatial structure of binding cavity, AcerASP2 could bind diverse odors with different structures, and the binding interaction could be easily to be promoted by the hydrogen bonds between odors with lysines in AcerASP2.

Key words: Apis cerana cerana , general odorant binding proteins , binding function , molecular docking , binding mode

[1]马德风, 梁诗魁. 中国蜜粉源植物及其利用. 北京: 中国农业出版社, 1993: 57-101.

Ma D F, Liang S K. Honey and Pollen Resource Plants in China. Beijing: China Agriculture Press, 1993: 57-101. (in Chinese)

[2]杨冠煌. 引入西方蜜蜂对中蜂的危害及生态影响. 昆虫学报, 2005, 48(3): 401-406.

Yang G H. Harm of introducing the western honeybee Apis mellifera L. to the Chinese honeybee Apis cerana F. and its ecological impact. Acta Entomologica Sinica, 2005, 48(3): 401-406. (in Chinese)

[3]Krieger J, Breer H. Olfactory reception in invertebrates. Science, 1999, 286(5440): 720-723.

[4]Pelosi P, Maida R. Odorant-binding proteins in insects. Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 1995, 111(3): 503-514.

[5]Pelosi P, Zhou J J, Ban L P, Calvello M. Soluble proteins in insect chemical communication. Cellular and Molecular Life Sciences, 2006, 63(14): 1658-1676.

[6]Nagnan-Le Meillour P, Huet J C, Maibeche M, Pernollet J C, Descoins C. Purification and characterization of multiple forms of odorant/pheromone binding proteins in the antennae of Mamestra brassicae (Noctuidae). Insect Biochemistry and Molecular Biology, 1996, 26(1): 59-67.

[7]Vogt R G, Prestwich G D, Lerner M R. Odorant-binding-protein subfamilies associate with distinct classes of olfactory receptor neurons in insects. Journal of Neurobiology, 1991, 22(1): 74-84.

[8]Forêt S, Maleszka R. Function and evolution of a gene family encoding odorant binding-like proteins in a social insect, the honey bee (Apis mellifera). Genome Research, 2006, 16(11): 1404-1413.

[9]Danty E, Arnold G, Huet J C, Huet D, Masson C, Pernollet J C. Separation, characterization and sexual heterogeneity of multiple putative odorant-binding proteins in the honeybee Apis mellifera L. (Hymenoptera: Apidea). Chemical Senses, 1998, 23(1): 83-91.

[10]Danty E, Michard-Vanhée C, Huet J C, Genecque E, Pernollet J C, Masson C. Biochemical characterization, molecular cloning and localization of a putative odorant-binding protein in the honey bee Apis mellifera L. (Hymenoptera: Apidea). FEBS Letters, 1997, 414(3): 595-598.

[11]Briand L, Nespoulous C, Huet J C, Takahashi M, Pernollet J C. Ligand binding and physico-chemical properties of ASP2, a recombinant odorant-binding protein from honeybee (Apis mellifera L.). European Journal of Biochemistry, 2001, 268(3): 752-760.

[12]Lescop E, Briand L, Pernollet J C, Guittet E. Structural basis of the broad specificity of a general odorant-binding protein from honeybee. Biochemistry, 2009, 48(11): 2431-2441.

[13]Lartigue A, Gruez A, Briand L, Blon F, Bezirard V, Walsh M, Pernollet J C, Tegoni M, Cambillau C. Sulfur single-wavelength anomalous diffraction crystal structure of a pheromone-binding protein from the honeybee Apis mellifera L. The Journal of Biological Chemistry, 2004, 279(6): 4459-4464.

[14]Lagarde A, Spinelli S, Tegoni M, He X, Field L, Zhou J J, Cambillau C. The crystal structure of odorant binding protein 7 from Anopheles gambiae exhibits an outstanding adaptability of its binding site. Journal of Molecular Biology, 2011, 414(3): 401-412.

[15]Kruse S W, Zhao R, Smith D P, Jones D N. Structure of a specific alcohol-binding site defined by the odorant binding protein LUSH from Drosophila melanogaster. Nature Structural Biology, 2003, 10(9): 694-700.

[16]李红亮, 聂文敏, 高其康, 程家安. 中华蜜蜂气味结合蛋白ASP2 cDNA的克隆及原核表达. 中国农业科学, 2008, 41(3): 933-938.

Li H L, Nie W M, Gao Q K, Cheng J A. Cloning of cDNA encoding odorant binding protein ASP2 in working bee’s antenna of Apis cerana cerana and its prokaryotic expression. Scientia Agricultura Sinica, 2008, 41(3): 933-938. (in Chinese)

[17]Li H L, Zhang Y L, Gao Q K, Cheng J A, Lou B G. Molecular identification of cDNA, immunolocalization, and expression of a putative odorant-binding protein from an Asian honeybee, Apis cerana cerana. Journal of Chemical Ecology, 2008, 34(12): 1593-1601.

[18]李红亮, 高其康, 程家安. 中华蜜蜂信息素结合蛋白ASP1 cDNA的克隆及时空表达. 昆虫学报, 2008, 51(7): 689-693.

Li H L, Gao Q K, Cheng J A. Cloning and spatio-temporal expression of cDNA encoding pheromone binding protein ASP1 in Apis cerana cerana (Hymenoptera: Apidae). Acta Entomologica Sinica, 2008, 51(7): 689-693. (in Chinese)

[19]李红亮, 倪翠侠, 姚瑞, 高其康, 商晗武. 中华蜜蜂化学感受蛋白基因Acer-CSP1克隆与表达特征分析. 昆虫学报, 2010, 53(9): 962-968.

Li H L, Ni C X, Yao R, Gao Q K, Shang H W. Molecular cloning, characterization, and expression pattern of chemosensory protein 1 gene (Acer-CSP1) in the Chinese honeybee, Apis cerana cerana (Hymenoptera: Apidae). Acta Entomologica Sinica, 2010, 53(9): 962-968. (in Chinese)

[20]Li H L, Lou B G, Cheng J A, Gao Q K. The chemosensory protein of Chinese honeybee, Apis cerana cerana: molecular cloning of cDNA, immunocytochemical localization and expression. Chinese Science Bulletin, 2007, 52(10): 1355-1364.

[21]Li H L, Zhang Y L, Wang H Y, Gao Q K, Cheng J A. Spatio-temporal expression analysis of two kinds of chemical communication-related proteins in the worker bee Apis cerana cerana Fabricius (Hymenoptera: Apidae). Chinese Journal of Agricultural Biotechnology, 2009, 6(2): 147-152.

[22]范成平, 商志才, 郭明, 俞庆森. 荧光法研究加潜沙星与牛乳铁蛋白的相互结合作用. 浙江大学学报: 理学版, 2005, 32(1): 62-65.

Fan C P, Shang Z C, Guo M, Yu Q C. Study on the interaction between gatifloxacin and bovine lactoferrin by fluorescence spectrophotometry. Journal of Zhejiang University: Science Edition, 2005, 32(1): 62-65. (in Chinese)

[23]李红亮, 张林雅, 倪翠侠, 商晗武. 中华蜜蜂化学感受蛋白AcerCSP3的配基结合功能分析. 昆虫学报, 2011, 54(3): 259-264.

Li H L, Zhang L Y, Ni C X, Shang H W. Molecular binding characterization with chemical ligands of a chemosensory protein AcerCSP3 in the Chinese honeybee, Apis cerana cerana Fabricius (Hymenoptera: Apidae). Acta Entomologica Sinica, 2011, 54(3): 259-264. (in Chinese)

[24]Gong Z J, Zhou W W, Yu H Z, Mao C G, Zhang C X, Cheng J A, Zhu Z R. Cloning, expression and functional analysis of a general odorant-binding protein 2 gene of the rice striped stem borer, Chilo suppressalis (Walker) (Lepidoptera: Pyralidae). Insect Molecular Biology, 2009, 18(3): 405-417.

[25]Arnold K, Bordoli L, Kopp J, Schwede T. The SWISS-MODEL workspace: a web-based environment for protein structure homology modelling. Bioinformatics, 2006, 22(2): 195-201.

[26]Deng S, Yin J, Zhong T, Cao Y, Li K. Function and immunocytochemical localization of two novel odorant-binding proteins in olfactory sensilla of the scarab beetle Holotrichia oblita Faldermann (Coleoptera: Scarabaeidae). Chemical Senses, 2012, 37(2): 141-150.

[27]Dani F R, Iovinella I, Felicioli A, Niccolini A, Calvello M A, Carucci M G, Qiao H, Pieraccini G, Turillazzi S, Moneti G, Pelosi P. Mapping the expression of soluble olfactory proteins in the honeybee. Journal of Proteome Research, 2010, 9(4): 1822-1833.
[1] QIU YiLei,WU Fan,ZHANG Li,LI HongLiang. Effects of Sublethal Doses of Imidacloprid on the Expression of Neurometabolic Genes in Apis cerana cerana [J]. Scientia Agricultura Sinica, 2022, 55(8): 1685-1694.
[2] LI GuiXiang,LI XiuHuan,HAO XinChang,LI ZhiWen,LIU Feng,LIU XiLi. Sensitivity of Corynespora cassiicola to Three Common Fungicides and Its Resistance to Fluopyram from Shandong Province [J]. Scientia Agricultura Sinica, 2022, 55(7): 1359-1370.
[3] ZHAO HuiTing,PENG Zhu,JIANG YuSuo,ZHAO ShuGuo,HUANG Li,DU YaLi,GUO LiNa. Expression and Binding Properties of Odorant Binding Protein AcerOBP7 in Apis cerana cerana [J]. Scientia Agricultura Sinica, 2022, 55(3): 613-624.
[4] ZHANG Li,ZHANG Nan,JIANG HuQiang,WU Fan,LI HongLiang. Molecular Cloning and Expression Pattern Analysis of NPC2 Gene Family of Apis cerana cerana [J]. Scientia Agricultura Sinica, 2022, 55(12): 2461-2471.
[5] FENG RuiRong,FU ZhongMin,DU Yu,ZHANG WenDe,FAN XiaoXue,WANG HaiPeng,WAN JieQi,ZHOU ZiYu,KANG YuXin,CHEN DaFu,GUO Rui,SHI PeiYing. Identification and Analysis of MicroRNAs in the Larval Gut of Apis cerana cerana [J]. Scientia Agricultura Sinica, 2022, 55(1): 208-218.
[6] XiaoHe LIU,GuiSheng QIU,ZhaoGuo TONG,HuaiJiang ZHANG,WenTao YAN,Qiang YUE,LiNa SUN. Ligands Binding Characteristics of Chemosensory Protein CsasCSP16 of Carposina sasakii [J]. Scientia Agricultura Sinica, 2021, 54(5): 945-958.
[7] QIN JianHui,LI JinQiao,ZHAO Xu,LI KeBin,CAO YaZhong,YIN Jiao. Expression, Purification and Functional Analysis of Odorant Binding Protein 11 (OBP11) in Anomala corpulenta [J]. Scientia Agricultura Sinica, 2021, 54(14): 3017-3028.
[8] LI ZuRen,LUO DingFeng,BAI HaoDong,XU JingJing,HAN JinCai,XU Qiang,WANG RuoZhong,BAI LianYang. Cloning and Expression Analysis of Light Harvesting Chlorophyll a/b Protein Gene CcLhca-J9 in Conyza canadensis [J]. Scientia Agricultura Sinica, 2021, 54(1): 86-94.
[9] CHEN WenFeng,WANG HongFang,LIU ZhenGuo,ZHANG WeiXing,CHI XuePeng,XU BaoHua. Recombinant Expression and Antimicrobial Activity of Apidaecin in Apis cerana cerana [J]. Scientia Agricultura Sinica, 2019, 52(4): 767-776.
[10] XU XiJian, GUO Rui, LUO Qun, XIONG CuiLing, LIANG Qin, ZHANG ChuanLian, ZHENG YanZhen, ZHANG ZhaoNan, HUANG ZhiJian, ZHANG Lu, LI WenDong, CHEN DaFu. De novo Transcriptome Assembly for Apis cerana cerana Larval Gut and Identification of SSR Molecular Markers [J]. Scientia Agricultura Sinica, 2017, 50(6): 1157-1166.
[11] TAN Jing, SONG XinMi, FU XiaoBin, TANG MingZhu, WU Fan, HUA QiYun, LI HongLiang. Functional Mode and Immunocytochemical Localization of Chemosensory Protein 1 (CSP1) in Apis cerana cerana [J]. Scientia Agricultura Sinica, 2017, 50(15): 3052-3062.
[12] CHEN DaFu, GUO Rui, XIONG CuiLing, LIANG qin, ZHENG YanZhen, XU XiJian, ZHANG ZhaoNan, HUANG ZhiJian, ZHANG Lu, WANG HongQuan, XIE YanLing, TONG XinYu. Transcriptome of Apis cerana cerana larval gut Under the Stress of Ascosphaera apis [J]. Scientia Agricultura Sinica, 2017, 50(13): 2614-2623.
[13] QIN Ming, WANG HongFang, LIU ZhenGuo, WANG Ying, WANG Shuai, CHI XuePeng, LIU ChunLei, ZHANG WeiXing, XU BaoHua. Comparison of Different Cold Resistance betweenApis cerana cerana and Apis mellifera ligustica [J]. Scientia Agricultura Sinica, 2017, 50(12): 2380-2388.
[14] ZHANG Li-zhen, WANG Zi-long, ZENG Zhi-jiang. Cloning and Expression of cAMP Response Element Binding Protein (CREB) Gene of Apis cerana cerana [J]. Scientia Agricultura Sinica, 2016, 49(8): 1599-1606.
[15] ZHANG Xiao, HU Xiao-dan, ZHONG Jian-feng, WU Ai-hua, XU Chong-xin, LIU Yuan, ZHANG Cun-zheng, XIE Ya-jing, LIU Xian-jin. Expression and Molecular Simulation of Alkaline Phosphatase Receptor of Plutella xyllostella [J]. Scientia Agricultura Sinica, 2016, 49(23): 4555-4565.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备09089781号-30
Copyright 2018 © Editorial office of Scientia Agricultura Sinica
Tel: 86-010-82106279    E-mail: zgnykx@mail.caas.net.cn
Supported by Beijing Magtech Co.Ltd