Functional Characteristics of a Novel Chemosensory Protein in the Cotton Bollworm Helicoverpa armigera (Hübner)

doi:10.1016/S2095-3119(13)60304-4

Journal of Integrative Agriculture ›› 2013, Vol. 12 ›› Issue (5): 853-861.DOI: 10.1016/S2095-3119(13)60304-4

Functional Characteristics of a Novel Chemosensory Protein in the Cotton Bollworm Helicoverpa armigera (Hübner)

ZHANG Tian-tao, WANG Wei-xuan, ZHANG Zi-ding, ZHANG Yong-jun , GUO Yu-yuan

1.State Key Laboratory for Biology of Plant Diseases and Insect Pests/Institute of Plant Protection, Chinese Academy of Agricultural
Sciences, Beijing 100193, P.R.China
2.State Key Laboratory of Agrobiotechnology/College of Biological Sciences, China Agricultural University, Beijing 100193, P.R.China

收稿日期:2012-09-09 出版日期:2013-05-01 发布日期:2013-05-01
通讯作者: Correspondence GUO Yu-yuan, Tel/Fax: +86-10-62894786; E-mail: yuyuanguo@hotmail.com; ZHANG Yong-jun, Tel: +86-10-62815929, E-mail: yjzhang@ippcaas.cn
作者简介:ZHANG Tian-tao, Mobile: 13718631910, E-mail: zhtiantao@163.com;
基金资助:
This work was supported by the National 973 Program of China (2012CB114104) and the National Natural Science Foundation of China (31171858).

Functional Characteristics of a Novel Chemosensory Protein in the Cotton Bollworm Helicoverpa armigera (Hübner)

ZHANG Tian-tao, WANG Wei-xuan, ZHANG Zi-ding, ZHANG Yong-jun , GUO Yu-yuan

1.State Key Laboratory for Biology of Plant Diseases and Insect Pests/Institute of Plant Protection, Chinese Academy of Agricultural
Sciences, Beijing 100193, P.R.China
2.State Key Laboratory of Agrobiotechnology/College of Biological Sciences, China Agricultural University, Beijing 100193, P.R.China

Received:2012-09-09 Online:2013-05-01 Published:2013-05-01
Contact: Correspondence GUO Yu-yuan, Tel/Fax: +86-10-62894786; E-mail: yuyuanguo@hotmail.com; ZHANG Yong-jun, Tel: +86-10-62815929, E-mail: yjzhang@ippcaas.cn
About author:ZHANG Tian-tao, Mobile: 13718631910, E-mail: zhtiantao@163.com;
Supported by:
This work was supported by the National 973 Program of China (2012CB114104) and the National Natural Science Foundation of China (31171858).

摘要/Abstract

摘要： A chemosensory protein named HarmCSP5 in cotton bollworm Helicoverpa armigera (Hübner) was obtained from antennal cDNA libraries and expressed in Escherichia coli. The real time quantitative PCR (RT-qPCR) results indicated that HarmCSP5 gene was mainly expressed in male and female antennae but also expressed in female legs and wings. Competitive binding assays were performed to test the binding affinity of recombinant HarmCSP5 to 60 odor molecules including some cotton volatiles. The resules showed that HarmCSP5 showed strong binding abilities to 4-ehtylbenzaldehyde and 3,4-dimethlbenz aldehyde, whereas methyl phenylacetate, 2-decanone, 1-pentanol, carvenol, isoborneol, nerolidol, 2- nonanone and ethyl heptanoate have relatively weak binding affinity. Moreover, the predicted 3D model of HarmCSP5 consists of six α-helices located among residues 33-38 (α1), 40-48 (α2), 62-72 (α3), 80-96 (α4), 98-108 (α5), and 116-119 (α6), two pairs of disulfide bridges Cys49-Cys55, Cys75-Cys78. The two amino acid residues, Ile94 and Trp101, may play crucial roles in HarmCSP5 binding with ligands and need further study for confirmation.

关键词: Helicoverpa armigera , chemosensory protein , expression characteristics , competitive binding assay , 3D structure model

Abstract: A chemosensory protein named HarmCSP5 in cotton bollworm Helicoverpa armigera (Hübner) was obtained from antennal cDNA libraries and expressed in Escherichia coli. The real time quantitative PCR (RT-qPCR) results indicated that HarmCSP5 gene was mainly expressed in male and female antennae but also expressed in female legs and wings. Competitive binding assays were performed to test the binding affinity of recombinant HarmCSP5 to 60 odor molecules including some cotton volatiles. The resules showed that HarmCSP5 showed strong binding abilities to 4-ehtylbenzaldehyde and 3,4-dimethlbenz aldehyde, whereas methyl phenylacetate, 2-decanone, 1-pentanol, carvenol, isoborneol, nerolidol, 2- nonanone and ethyl heptanoate have relatively weak binding affinity. Moreover, the predicted 3D model of HarmCSP5 consists of six α-helices located among residues 33-38 (α1), 40-48 (α2), 62-72 (α3), 80-96 (α4), 98-108 (α5), and 116-119 (α6), two pairs of disulfide bridges Cys49-Cys55, Cys75-Cys78. The two amino acid residues, Ile94 and Trp101, may play crucial roles in HarmCSP5 binding with ligands and need further study for confirmation.

Key words: Helicoverpa armigera , chemosensory protein , expression characteristics , competitive binding assay , 3D structure model

ZHANG Tian-tao, WANG Wei-xuan, ZHANG Zi-ding, ZHANG Yong-jun , GUO Yu-yuan. Functional Characteristics of a Novel Chemosensory Protein in the Cotton Bollworm Helicoverpa armigera (Hübner)[J]. Journal of Integrative Agriculture, 2013, 12(5): 853-861.

参考文献

[1]Angeli S, Ceron F, Scaloni A, Monti M, Monteforti G,Minnocci A, Petacchi R, Pelosi P. 1999. Purification,s t r u c t u r a l c h a r a c t e r i z a t i o n , c l o n i n g a n dimmunocytochemical localization of chemoreceptionproteins from Schistocerca gregaria. European Journalof Biochemistry, 262, 745-754

[2]Ban L, Scaloni A, Brandazza A, Angeli S, Zhang L, Yan Y,Pelosi P. 2003. Chemosensory proteins of Locustamigratoria. Insect Molecular Biology, 12, 125-134

[3]Ban L, Zhang L, Yan Y, Pelosi P. 2002. Binding propertiesof a locust’s chemosensory protein. Biochemical andBiophysical Research Communications, 293, 50-54

[4]Benton R, Vannice K S, Gomez-Diaz C, Vosshall L B. 2009.Variant ionotropic glutamate receptors aschemosensory receptors in Drosophila. Cell, 136, 149-162

[5]Breer H, Raming K, Krieger J. 1994. Signal recognition andtransduction in olfactory neurons. Biochimica etBiophysica Acta, 1224, 277-287

[6]Briand L, Nespoulous C, Huet J C, Takahashi M, PernolletJ C. 2001. Ligand binding and physico-chemicalproperties of ASP2, a recombinant odorant-bindingprotein from honeybee (Apis mellifera L.). EuropeanJournal of Biochemistry, 268, 752-760

[7]Campanacci V, Lartigue A, Hallberg B M, Jones T A, Giudici-Orticoni M T, Tegoni M, Cambillau C. 2003. Mothchemosensory protein exhibits drastic conformationalchanges and cooperativity on ligand binding.Proceedings of the National Academy of Sciences ofthe United States of America, 100, 5069-5074

[8]Celorio-Mancera M P, Sundmalm S M, Vogel H, RutishauserD, Ytterberg A J, Zubarev R A, Janz N. 2012.Chemosensory proteins, major salivary factors incaterpillar mandibular glands. Insect Biochemistry andMolecular Biology, 42, 796-805

[9]Field L, Pickett J, Wadhams L. 2000. Molecular studies ininsect olfaction. Insect Molecular Biology, 9, 545-551

[10]Ginalski K, Elofsson A, Fischer D, Rychlewski L. 2003. 3DJury:a simple approach to improve protein structurepredictions. Bioinformatics, 19, 1015-1018

[11]Gong D, Zhang H, Zhao P, Lin Y, Xia Q, Xiang Z. 2007.Identification and expression pattern of thechemosensory protein gene family in the silkworm,Bombyx mori. Insect Biochemistry and MolecularBiology, 37, 266-277

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

[13]Guo W, Wang X, Ma Z, Xue L, Han J, Yu D, Kang L. 2011.CSP and takeout genes modulate the switch betweenattraction and repulsion during behavioral phase changein the migratory locust. PLoS Genetics, 7, e1001291.

[14]Hua J F, Zhang S, Cui J J, Wang D J, Wang C Y, Luo J Y, LvL M. 2012. Identification and binding characterizationof three odorant binding proteins and onechemosensory protein from Apolygus lucorum (Meyer-Dur). Journal of Chemical Ecology, 38, 1163-1170

[15]Jacquin-Joly E, Vogt R G, Francois M C, Nagnan-Le MeillourP. 2001. Functional and expression pattern analysis ofchemosensory proteins expressed in antennae andpheromonal gland of Mamestra brassicae. ChemicalSenses, 26, 833-844

[16]Jiang Q Y, Wang W X, Zhang Z, Zhang L. 2009. Bindingspecificity of locust odorant binding protein and itskey binding site for initial recognition of alcohols. InsectBiochemistry and Molecular Biology, 39, 440-447

[17]Kaissling K E, Strausfeld C Z, Rumbo E R. 1987. Adaptationprocesses in insect olfactory receptors. Annals of theNew York Academy of Sciences, 510, 104-112

[18]Kitabayashi A N, Arai T, Kubo T, Natori S. 1998. Molecularcloning of cDNA for p10, a novel protein that increasesin the regenerating legs of Periplaneta americana(American cockroach). Insect Biochemistry andMolecular Biology, 28, 785-790

[19]Kruger N J. 2002. The Bradford method for proteinquantitation. The Protein Protocols Handbook. 2nded. Humana Press Inc., Totowa, NJ. pp. 15-21

[20]Lartigue A, Campanacci V, Roussel A, Larsson A M, JonesT A, Tegoni M, Cambillau C. 2002. X-ray structure andligand binding study of a moth chemosensory protein.Journal of Biological Chemistry, 277, 32094-32098

[21]Liu R, He X, Lehane S, Lehane M, Hertz-Fowler C, BerrimanM, Field L M, Zhou J J. 2012. Expression ofchemosensory proteins in the tsetse fly Glossinamorsitans morsitans is related to female host-seekingbehaviour. Insect Molecular Biology, 21, 41-48

[22]Livak K J, Schmittgen T D. 2001. Analysis of relative geneexpression data using real-time quantitative PCR andthe 2-儮儮CT method. Methods, 25, 402-408

[23]Luthy R, Bowie J U, Eisenberg D. 1992. Assessment ofprotein models with three-dimensional profiles. Nature,356, 83-85

[24]Mckenna M P, Hekmat-Scafe D S, Gaines P. 1994. PutativeDrosophila pheromone-binding proteins expressed ina subregion of the olfactory system. The Journal ofBiological Chemistry, 269, 16340-16347

[25]Oduol F, Xu J, Niare O, Natarajan R, Vernick K D. 2000.Genes identified by an expression screen of the vectormosquito Anopheles gambiae display differentialmolecular immune response to malaria parasites andbacteria. Proceedings of the National Academy ofSciences of the United States of America, 97, 11397-11402

[26]Pelosi P, Zhou J, Ban L, Calvello M. 2006. Soluble proteinsin insect chemical communication. Cellular andMolecular Life Sciences, 63, 1658-1676

[27]Permyakov E A, Zhang Y, Dong X, Liu J, Hu M, Zhong G,Geng P, Yi X. 2012. Molecular cloning, expression andmolecular modeling of chemosensory protein fromSpodoptera litura and its binding properties withrhodojaponin III. PLoS ONE, 7, e47611.Picimbon J F, Dietrich K, Krieger J, Breer H. 2001. Identityand expression pattern of chemosensory proteins inHeliothis virescens (Lepidoptera, Noctuidae). InsectBiochemistry and Molecular Biology, 31, 1173-1181

[28]Sabatier L, Jouanguy E, Dostert C, Zachary D, Dimarcq J L,Bulet P, Imler J L. 2003. Pherokine-2 and -3

[29]EuropeanJournal of Biochemistry, 270, 3398-3407

[30]Sali A, Blundell T L. 1993. Comparative protein modellingby satisfaction of spatial restraints. Journal ofMolecular Biology, 234, 779-815

[31]Stathopoulos A, van Drenth M, Erives A, Markstein M,Levine M. 2002. Whole-genome analysis of dorsalventralpatterning in the Drosophila embryo. Cell, 111,687-701

[32]Tomaselli S, Crescenzi O, Sanfelice D, Wechselberger R,Angeli S, Scaloni A, Boelens R, Tancredi T, Pelosi P,Picone D. 2006. Solution structure of a chemosensoryprotein from the desert locust Schistocerca gregaria.Biochemistry, 45, 10606-10613

[33]Vogt R G. 2002. Odorant binding protein homologues ofthe malaria mosquito Anopheles gambiae possibleorthologues of the OS-E and OS-F OBPs of Drosophilamelanogaster. Journal of Chemical Ecology, 28, 2371-2376

[34]Vogt R G, Miller N E, Litvack R, Fandino R A, Sparks J,Staples J, Friedman R, Dickens J C. 2009. The insectSNMP gene family. Insect Biochemistry and MolecularBiology, 39, 448-456

[35]Vogt R G, Riddiford L M. 1981. Pheromone binding andinactivation by moth antennae. Nature, 293, 161-163

[36]Vogt R G, Riddiford L M, Prestwich G D. 1985. Kineticproperties of a pheromone degrading enzyme: thesensillar esterase of Antheraea polyphemus.Proceedings of the National Academy of Sciences ofthe United States of America, 82, 8827-8831

[37]Wanner K W, Willis L G, Theilmann D A, Isman M B, FengQ, Plettner E. 2004. Analysis of the insect os-d-like genefamily. Journal of Chemical Ecology, 30, 889-911

[38]Xu Y L, He P, Zhang L, Fang S Q, Dong S L, Zhang Y J, LiF. 2009. Large-scale identification of odorant-bindingproteins and chemosensory proteins from expressedsequence tags in insects. BMC Genomics, 10, 632.Yu H L, Zhang Y J, Pan W L, Guo Y Y, Gao X W. 2007.Identification of volatiles from field cotton plant underdifferent induction treatments. Chinese Journal ofApplied Ecology, 18, 859-864

[39](in Chinese)Zhang T T, Gu S H, Wu K M, Zhang Y J, Guo Y Y. 2011.Construction and analysis of cDNA libraries from theantennae of male and female cotton bollwormsHelicoverpa armigera (Hübner) and expressionanalysis of putative odorant-binding protein genes.B i o c h e m i c a l a n d B i o p h y s i c a l R e s e a r c hCommunications, 407, 393-399

[40]Zhang T T, Wang W X, Zhang Z D, Zhang Y J, Guo Y Y.2012. Structure, binding characteristics, and 3D modelprediction of a newly identified odorant-binding proteinfrom the cotton bollworm, Helicoverpa armigera(Hübner). Journal of Integrative Agriculture, 11, 430-438

[41]Zhou J J. 2010. Odorant-binding proteins in insects.Vitamins & Hormones, 83, 241-272.

Functional Characteristics of a Novel Chemosensory Protein in the Cotton Bollworm Helicoverpa armigera (Hübner)

Functional Characteristics of a Novel Chemosensory Protein in the Cotton Bollworm Helicoverpa armigera (Hübner)

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