昆蟲氣味結(jié)合蛋白研究進展

張治科，張 燁，吳圣勇，雷仲仁, 4*

昆蟲氣味結(jié)合蛋白研究進展

張治科1*，張 燁2，吳圣勇3，雷仲仁3, 4*

(1.寧夏農(nóng)林科學(xué)院植物保護研究所，寧夏植物病蟲害防治重點實驗室，銀川 750002；2.山西省農(nóng)業(yè)科學(xué)院植物保護研究所，太原 030031；3.中國農(nóng)業(yè)科學(xué)院植物保護研究所，植物病蟲害生物學(xué)國家重點實驗室，北京 100193；4.閩臺特色作物病蟲生態(tài)防控協(xié)同創(chuàng)新中心，福州 350002))

嗅覺在昆蟲生命活動中起著重要的作用，氣味結(jié)合蛋白(odorant binding proteins, OBPs)是昆蟲嗅覺系統(tǒng)中發(fā)揮重要作用的蛋白之一，近年隨著基因組學(xué)、轉(zhuǎn)錄組學(xué)的快速發(fā)展，越來越多的昆蟲OBPs基因陸續(xù)被鑒定出來，部分OBPs的功能也逐步被證實。本文作者針對OBPs的種類、結(jié)構(gòu)特征、表達分布、三維結(jié)構(gòu)以及生理功能等方面進行了綜述，為更多昆蟲OBPs基因的鑒定及其功能研究提供參考，也為進一步揭示昆蟲-環(huán)境間的化學(xué)通訊機理以及開辟害蟲新的防治策略奠定基礎(chǔ)。

昆蟲嗅覺；氣味結(jié)合蛋白；結(jié)構(gòu)特征；表達分布；生理功能

昆蟲在長期進化過程中演化出復(fù)雜的嗅覺、味覺、觸覺等信號感受機制，其中嗅覺在昆蟲寄主選擇、交尾、覓食、忌避及信息傳遞等生命活動中發(fā)揮重要作用。氣味結(jié)合蛋白(odorant binding proteins, OBPs)是昆蟲嗅覺系統(tǒng)中發(fā)揮重要作用的蛋白之一，主要存在于觸角嗅覺感受器胞外空間的淋巴液內(nèi)，能夠結(jié)合并運送脂溶性氣味分子通過水溶性淋巴液到達嗅覺神經(jīng)元樹突，激活樹突膜上的嗅覺受體引發(fā)第二信使反應(yīng)(Mombaerts, 1999)。由于昆蟲觸角上存在大量表皮具有微孔的嗅覺相關(guān)感器，通常認為親脂性的氣味分子由這些微孔進入到感器淋巴液中，被OBPs識別后結(jié)合，然后運轉(zhuǎn)到氣味受體(olfactory receptors, Ors)，啟動G蛋白偶聯(lián)信號途徑，嗅覺受體神經(jīng)元上的Ors檢測到氣味物質(zhì)并被識別和激活，引起膜電位發(fā)生變化并刺激神經(jīng)，經(jīng)軸突傳到中樞神經(jīng)系統(tǒng)，大腦整合后發(fā)出指令，從而產(chǎn)生相關(guān)嗅覺行為反應(yīng)(Krieger and Breer, 2003)，隨后這些氣味分子被感器淋巴腔中的氣味降解酶(odorant degrading enzymes, ODEs)或者感器支持細胞中的多種酶降解(Vogtetal., 2005)，將氣味物質(zhì)迅速分解，阻止氣味物質(zhì)與受體持續(xù)結(jié)合，從而保持受體活性(Vogt and Riddiford, 1981)。

1 OBPs種類及鑒定

自從Vogt等(1981)采用同位素標記的方法，起始在雄性多音天蠶蛾Antheraeapolyphemus的觸角中發(fā)現(xiàn)了一個信息素結(jié)合蛋白(pheromone binding proteins，PBPs)，其分子量16 kDa、等電點4.7、能夠特異性結(jié)合雌性信息素、大量存在于毛形感器淋巴液中的可溶性蛋白，這是首次發(fā)現(xiàn)的一個昆蟲OBP，開啟了OBPs的研究，并在近些年來一直是昆蟲化學(xué)生態(tài)學(xué)研究的熱點。OBPs在昆蟲觸角感器淋巴液中濃度非常高，進一步細分為PBPs和普通OBPs(general odorant binding proteins，GOBPs)，其中GOBPs又分為普通OBPsⅠ(general odorant binding proteinsⅠ，GOBPⅠ)和普通OBPsII(general odorant binding proteins II，GOBPII)(任珍珍等，2010)；還有一類稱為觸角特異蛋白(antenna special proteins，ASPs)，最初在西方蜜蜂ApismelliferaL.中發(fā)現(xiàn)，包括觸角特異蛋白ASP1和ASP2(Dantyetal., 1998)。PBPs主要感受雌蟲釋放的性信息素組分，分布于毛型感器中，GOBPs主要感受環(huán)境中植物揮發(fā)性化學(xué)物質(zhì)等環(huán)境信號氣味物質(zhì)，分布于錐型感器中。

近年來，越來越多的昆蟲OBPs陸續(xù)被鑒定出來，如鱗翅目(曹馨月等，2015；李文海等，2015)、半翅目(Vandermotenetal., 2011)、雙翅目(陳玲等，2013)、直翅目(Yuetal., 2009)、鞘翅目(Juetal., 2012)、膜翅目(吉挺等，2014)等。再加上基因組學(xué)、轉(zhuǎn)錄組學(xué)的發(fā)展速度飛快，研究人員利用這些技術(shù)手段，陸續(xù)發(fā)現(xiàn)了較多個OBPs，如在膜翅目昆蟲意大利蜜蜂A.melliferaligustica中鑒定了21個OBPs(Forêt and Maleszka, 2006; Zhaoetal., 2013)；在雙翅目昆蟲黑腹果蠅Drosophilamelanogaster基因組中鑒定了近60個OBPs(Matsuoetal., 2007)，在中華按蚊Anophelessinensis中鑒定了64個OBPs(Xiuetal., 2016)；在鞘翅目昆蟲暗黑鰓金龜Holotrichiaparallela中鑒定了25個OBPs(Juetal., 2014)；在直翅目昆蟲亞洲小車蝗Oedaleusasiaticus觸角中鑒定了15個OBPs(張碩，2015)；在鱗翅目昆蟲棉鈴蟲的觸角轉(zhuǎn)錄組中鑒定了26個OBPs(Liuetal., 2012)，在家蠶Bombyxmori的基因組中，發(fā)現(xiàn)了40多個OBPs(Gongetal., 2009a)；在同翅目昆蟲中也鑒定了較多個OBPs，如Zhou等(2010)在豌豆蚜中發(fā)現(xiàn)了十幾個OBPs。OBPs蛋白序列相似性差異較大，有的序列相似性能到達90%以上，有的卻小于10%，尤其是不同目昆蟲或不同種昆蟲OBPs間。

2 昆蟲OBPs氨基酸序列特征

昆蟲OBPs由位于嗅覺感器旁邊的支持細胞合成，然后釋放到感器的淋巴液中，是一類小分子(14-17 kDa)水溶性、偏酸性(pH5.0左右)的球狀蛋白(Pelosietal., 2006)，能與進入到觸角內(nèi)的脂溶性的氣味物質(zhì)結(jié)合并承擔(dān)運送和卸載信息化合物的角色，是昆蟲專一性識別環(huán)境氣味物質(zhì)的第一步生化反應(yīng)(Vogt and Riddiford, 1981)，對昆蟲與外界進行信息交流起到重要意義(紀萍等，2013)。昆蟲OBPs蛋白通常由135-220個氨基酸組成，不同昆蟲種間序列相似性較低，但典型OBPs結(jié)構(gòu)中均存在六個保守的半胱氨酸位點(Krieger and Breer，1999)，且第2和第3個Cys之間間隔3個氨基酸，第5和第6個Cys之間間隔8個氨基酸；蛋白三維結(jié)構(gòu)中成對出現(xiàn)的3個二硫鍵由這6個保守半胱氨酸形成的，對蛋白三維結(jié)構(gòu)起到穩(wěn)固作用(Tegonietal., 2004)，如家蠶BmorPBP中Csy19-Csy54、Csy50-Csy108和Csy97-Csy117間形成的3個二硫鍵分別連接α螺旋1和3、3和6，5和6(Sandleretal., 2000)。還有一類非典型的OBPs，含有多于或少于6個保守的半胱氨酸位點，分別稱為Plus-C OBP(Zhouetal., 2004)和Minus-C OBP(Spinellietal., 2012)。通常研究認為，OBPs的主要功能是首先識別并溶解環(huán)境中的脂溶性氣味分子，然后承載這些氣味分子、通過感器淋巴液、到達Ors，激活了整個嗅覺信號的轉(zhuǎn)導(dǎo)過程(Krieger, 1999)。

3 OBPs表達分布特征

起初研究者們發(fā)現(xiàn)，昆蟲OBPs僅在觸角中表達，如家蠶BmorOBPs(Kriegeretal., 1996)、亞洲玉米螟Ostriniafurnacalis的OfurPBP3(Allen and Wanner, 2011)、棉鈴蟲的HarmPBPs(Guoetal., 2012)和HarmOBP2(Wangetal., 2003)、小菜蛾P(guān)lutellaxylostella的PxylPBP1(Zhangetal., 2009)、小地老虎Agrotisipsilon的AipsPBPl-3(Huaetal., 2012)、水稻二化螟Chilosuppressalis的CsupOBP2(Gongetal., 2009b)等。

后來大量的研究證據(jù)表明，有些OBPs不僅在觸角中表達，也在昆蟲的頭、胸、腹、足、翅、下顎須、喙、口器、性腺、精囊、氣門等其它組織部位中表達(Lietal., 2008；Vogeletal., 2010；Huaetal., 2012；Yinetal., 2012；陳玲等，2013；魏丹等，2013；Zhuetal., 2013；吉挺等，2014；秦贈等，2014；宋月芹等，2014；趙雪等，2014)。

有的OBPs在雌雄蟲體中的表達量和部位也有較大差異，如水稻二化螟Minus-C CsupOBP1在雄蟲觸角內(nèi)的表達量顯著高于雌蟲觸角(魏丹等，2013)，小菜蛾的PxylPBPs在雄性成蟲的足以及雌性成蟲的生殖器官中表達(Sunetal., 2013)，斜紋夜蛾的 SlitPBPl和SlitPBP2在表達于雄性成蟲中表達十分高、斜紋夜蛾的SlitPBP3在雌性成蟲中大量表達(Liuetal., 2013)，小地老虎PBP1、PBP2和PBP3在雄性觸角中表達，在味覺器官喙和下唇須中也有少量的表達(Guetal., 2013)，在雌性成蟲的腹部末端CpomGOBP2-3有表達(Garczynskietal., 2012)。

有的OBPs在蚜蟲有翅和無翅期的表達量也存在差異，如茄無網(wǎng)蚜AcyrthosiphonsolaniOBPs OBP7在無翅成蚜?xí)r表達量顯著高于有翅蚜(趙雪等，2014)，推測該蛋白在無翅成蚜的覓食等嗅覺行為活動中發(fā)揮著重要作用。

近年來，大量的免疫組織化學(xué)和原位雜交實驗表明，GOBP主要在錐形感器中表達，PBPs通常在毛形感器淋巴液中表達(Forstneretal., 2009; Guetal., 2013)。有些OBPs在毛形感器和錐形感器淋巴液中均有表達(Guetal., 2011)。

4 OBPs三維結(jié)構(gòu)研究

三維結(jié)構(gòu)能夠更加直觀的闡明、推測OBPs的功能，對深入研究OBPs的功能起到不可或缺的作用。目前，NCBI數(shù)據(jù)庫中已提交數(shù)千個PBPs序列，而PDB數(shù)據(jù)庫中提交的OBPs晶體結(jié)構(gòu)并不多。目前比較經(jīng)典的研究昆蟲OBPs三維結(jié)構(gòu)的兩種方法是X射線衍射(X-ray diffraction spectroscopy)和核磁共振技術(shù)(nuclear magnetic resonance, NMR)。昆蟲OBPs三維結(jié)構(gòu)第一個獲得解析的是家蠶的BmorPBP(Dambergeretal., 2000)，該蛋白結(jié)合口袋是由螺旋α1、α4、α5、α6組成，α3覆蓋在結(jié)合口袋的另一端，結(jié)合口袋內(nèi)部結(jié)合有性信息素蠶蛾醇(Sandleretal., 2000)。研究發(fā)現(xiàn)，pH值對PBPs的三維結(jié)構(gòu)影響較大，家蠶復(fù)合物BmorPBP-bombykol的三維結(jié)構(gòu)會因觸角淋巴液pH的不同而有變化，如在高PH條件下表現(xiàn)出親和性，低PH值條件下沒有親和性(Wojtasek and Leal，1999)，這種現(xiàn)象在重建的BmorPBP1和天然蛋白中均存在，推測蛋白PH值從高向低轉(zhuǎn)換是與生理相關(guān)的。隨后研究證明在高、低兩種PH值條件下，性信息素分子與家蠶BmorPBP的C末端折疊形成的α-螺旋共同競爭BmorPBP結(jié)合腔，當在pH較高時，BmorPBP蛋白的C末端會在蛋白的表面上折疊，然后開始結(jié)合性信息素，蠶蛾醇占據(jù)結(jié)合腔；低pH條件下BmorPBP1的C末端折疊形成一個α-螺旋占據(jù)結(jié)合腔，此時釋放性信息素(Sandleretal., 2000)。由于樹突表面的PH值較低(Leal, 2005a)，當蠶蛾醇或BmorPBP到達Ors時信息素分子便從結(jié)合腔內(nèi)釋放出來。

隨后又有果蠅的LUSH(Kruseetal., 2003)、蜚蠊Leucophaeamaderae的LmadPBP(Lartigueetal., 2003)、多音天蠶蛾的ApolPBP(Mohantyetal., 2004)、意大利蜜蜂的AmelPBP(Lartigueetal., 2004)、AmelGOBP(Lescopetal., 2009)和C-minus OBP AmelOBP14(Spinellietal., 2012)、岡比亞按蚊Anophelesgambiae的AgamOBP1(Wogulisetal., 2006)、家蠶的BmorGOBP2(Zhouetal., 2009)等的三維結(jié)構(gòu)陸續(xù)獲得了解析。

目前采用同源模建的方法，尤其在序列一致性較高的情況下，能夠較為準確的預(yù)測OBPs的三維結(jié)構(gòu)，促進了OBPs功能的進一步研究。如利用同源模建法構(gòu)建棉紅鈴蟲Pectinophoragossypiella的PBP三維結(jié)構(gòu)，并采用Errat、Verify_3D、Procheck、ProSa2003等程序的評價結(jié)果具有很高的可靠性，顯示棉紅鈴蟲PBPs結(jié)構(gòu)主要由6個α-螺旋和連接這些螺旋的回折構(gòu)成，底物結(jié)合口袋成錐形，6個保守的半胱氨酸形成3個二硫鍵，對蛋白結(jié)構(gòu)起到穩(wěn)定作用(孫浩等，2013)；中華蜜蜂Apisceranacerana體外重組蛋白AcerASP2與氣味信息的結(jié)合模式和機理也通過同源建模和分子對接進行解析，顯示4-烯丙基藜蘆醚絕大部分位于該重組蛋白預(yù)測的1個狹長口袋狀的疏水性結(jié)合腔內(nèi)，并與Lys74產(chǎn)生2個氫鍵(李紅亮等，2013)；花絨寄甲Dastarcushelophoroides的DhelOBP21通過配體結(jié)合實驗和分子對接，疏水相互作用比氫鍵相互作用更為明顯，盡管形成氫鍵的相互作用可以預(yù)測一些結(jié)合復(fù)合物，疏水相互作用更大程度的影響著疏水性結(jié)合腔的變化，配體的取向通過影響疏水相互作用而影響結(jié)合(Lietal., 2015)；信息化合物誘烯醇與蘋果蠹蛾Cydiapomonella的CpomPBP2和BmorPBP對接的構(gòu)象疊加研究表明， CpomPBP2結(jié)合袋內(nèi)整個化合物的準確對接(Tianetal., 2016)；基于靶標蛋白岡比亞按蚊AgamOBP1結(jié)合口袋特征，結(jié)合所構(gòu)建的?；哙ゎ惢锶S定量構(gòu)效關(guān)系模型，闡明了?；哙ゎ惢衔锝Y(jié)構(gòu)與驅(qū)避活性的關(guān)系(陳文雅等，2013)，等。

昆蟲OBPs的結(jié)構(gòu)及其結(jié)合特性研究表明，6個α-螺旋構(gòu)成的OBP的三維結(jié)構(gòu)的結(jié)合腔，很有可能就是與外界氣味分子結(jié)合的關(guān)鍵部位，而且氣味分子進出結(jié)合腔會與一些因子密切相關(guān)，Leal(2005b)研究報道如氣味分子的構(gòu)象、觸角感器淋巴液的pH值等。Sandler等(2000)報道在識別氣味分子時，位于蛋白結(jié)合腔開口處或內(nèi)部的某些親水性氨基酸殘基發(fā)揮重要角色，除此，OBPs結(jié)合化學(xué)氣味分子和釋放化學(xué)氣味分子時，都會經(jīng)歷pH致使的一些構(gòu)象變化，Xu等(2010)研究表明，當在酸性條件下，位于PBP C鏈末端的氨基酸會額外構(gòu)成1個a-螺旋，同時占據(jù)了結(jié)合腔的內(nèi)部，擠出了氣味分子，在中性條件下，氣味分子再次進入結(jié)合腔，由于C鏈末端又會變得疏松，這種現(xiàn)象與Kowcun等(2001)報道的位于嗅覺神經(jīng)元樹突膜附近的淋巴液通常情況下會偏酸性以及昆蟲感器附近的淋巴液普遍認為呈中性的觀點相一致。

5 OBPs生理功能

有關(guān)OBPs在昆蟲嗅覺識別中具體功能的假說不斷被提出，目前普遍認為OBPs可能具有的功能(趙紅霞等，2015)：(1)具有外周濾器作用。在氣味分子識別過程中選擇性結(jié)合某些氣味分子，同時在氣味分子濃度過高時OBPs可降低其濃度，以免降低受體靈敏度，如PBPs對非信息素分子具有過濾作用。(2)OBPs可以通過親水性的淋巴液作為溶劑和運輸氣味物質(zhì)的載體。被認為能夠溶解進入感器淋巴液中的氣味分子，并且能夠運輸一些脂溶性的氣味分子。(3)OBPs幫助氣味分子到達嗅覺受體蛋白，有利于信號迅速傳導(dǎo)。(4)降解氣味分子，使氣味分子失活。OBPs能夠?qū)馕斗肿舆\輸?shù)叫嵊X受體，然后迅速激活嗅覺受體后，又能夠迅速降解氣味分子。(5)起到清除作用，能夠及時清除結(jié)合在受體上的各種信息素，保持受體的活性。(6)OBPs還來自于昆蟲觸角以外的其它不同組織部位，意味著OBPs很可能還扮演其它未被揭示的重要功能。

截至目前科學(xué)家們能夠確認的是首先識別并結(jié)合親脂性氣味分子，然后運輸這些親脂性氣味分子，穿過感器淋巴液，最后到達目的地—嗅覺受體，有關(guān)其作用過程，有人研究認為OBP到達膜結(jié)合受體后釋放配體，然后配體激活嗅覺受體(Maoetal., 2010)；也有人認為OBP與氣味分子結(jié)合的復(fù)合物共同激活膜結(jié)合受體(Laughlinetal., 2008)。當然，OBP在昆蟲體內(nèi)其他組織中也有表達，可能還發(fā)揮著其他重要的功能(Leal, 2013)。OBP可以單獨、或與自身、或與其它OBP形成二聚體后與氣味分子結(jié)合，如岡比亞按蚊OBP1和OBP4可形成二聚體共表達(Schultzeetal., 2012)。

在20世紀80-90年代主要采用同位素標記性信息素，再經(jīng)聚丙烯凝膠電泳、轉(zhuǎn)印和放射自顯影技術(shù)檢測結(jié)果的方法對多音天蠶蛾、舞毒蛾、煙草天蛾、甘藍夜蛾等鱗翅目昆蟲進行氣味分子結(jié)合蛋白的功能研究(Lietal., 1997)。這種方法的缺點是不夠簡便、不安全并且實驗結(jié)果不穩(wěn)定。測定蛋白結(jié)合外界的氣味分子的方法有幾種，如Gu等(2011)采用的熒光競爭結(jié)合實驗，Zhou等(2009)采用的冷結(jié)合實驗，還有He等(2010)采用的雙相結(jié)合實驗。在這些方法當中，熒光競爭結(jié)合實驗被用得最為廣泛(Zhongetal., 2012)，該方法利用氣味分子與可發(fā)射熒光的高效特異探針和目的蛋白的競爭結(jié)合，能夠測定OBP與氣味分子特異性結(jié)合的強弱，可篩選出專一性的氣味物質(zhì)或性信息素，且簡便、安全、可靠，該方法已被廣泛應(yīng)用到OBP與氣味分子結(jié)合能力的測試中(Gongetal., 2009b；Yuetal., 2009；孫紅巖等，2011；陳玲等，2013；李紅亮等，2013；魏丹等，2013；宋月芹等，2014)。

6 展望

昆蟲嗅覺系統(tǒng)是一個高度專一、極其靈敏的化學(xué)檢測器，可以識別環(huán)境中的特異性化學(xué)氣味分子并直接接觸這些信號物質(zhì)進行覓食、趨避、尋偶和選擇產(chǎn)卵場所等生命活動(Schneider, 1969)，對昆蟲適應(yīng)環(huán)境和種群繁衍具有重要的生物學(xué)意義。探索、闡明昆蟲與環(huán)境之間信息聯(lián)系的本質(zhì)規(guī)律，并運用此規(guī)律研究開發(fā)害蟲產(chǎn)卵忌避劑和益蟲利用新技術(shù)等，一直是國內(nèi)外化學(xué)生態(tài)學(xué)研究的熱點和目標之一，是生物間化學(xué)通訊機制研究的主要內(nèi)容(Pelosietal., 2006)。

研究昆蟲嗅覺相關(guān)蛋白是闡明昆蟲嗅覺機理的基礎(chǔ)，OBPs是參與昆蟲嗅覺系統(tǒng)活動的重要成員之一，主要參與外界氣味物質(zhì)的嗅覺識別和傳遞過程。已報道的OBPs多源自鱗翅目、半翅目、雙翅目、等翅目、直翅目、鞘翅目、膜翅目等，還有許多目昆蟲OBPs至今未有鑒定。通過學(xué)者們不斷探索研究，陸續(xù)明確了OBPs的一些生理生化特征、證實了OBPs的某些重要生理功能，如對氣味物質(zhì)的識別、運輸、降解，以及OBPs與具體氣味或信息化合物的分子對接機理等，但OBPs也許還發(fā)揮著其它未被發(fā)現(xiàn)或證實的重要功能，尤其蛋白互做功能研究任重道遠、意義重大，尚需進一步深入研究，才能確切闡明OBPs在昆蟲行為中所發(fā)揮的重要功能。因此，在今后的工作中，仍需加大對更多目昆蟲OBPs的鑒定，加大對OBPs單獨及互做下所發(fā)揮的功能深入研究，更好的明確昆蟲寄主選擇、驅(qū)避、產(chǎn)卵、求偶等方面的生化和分子機制、揭示害蟲行為反應(yīng)的本質(zhì)原因、闡明昆蟲的嗅覺感受機理、解析害蟲-環(huán)境-寄主之間的化學(xué)通訊奠定基礎(chǔ)，從而為保護和利用有益昆蟲、調(diào)控昆蟲行為、研發(fā)環(huán)境友好型引誘劑或驅(qū)避劑等害蟲防控新策略提供依據(jù)。

References)

Allen JE, Wanner KW.Asian com borer pheromone binding protein 3, a candidate for evolving specificity to the 12-tetradecenyl acetate sex pheromone [J].InsectBiochemistryandMolecularBiology, 2011, 41: 141-149.

Cao XY, Huang YY, Zhao X,etal.Cloning and prokaryotic expression ofCydiapomonellapheromone binding proteinⅡ gene (CpomPBP2) [J].JournalofNorthwestA&FUniversity(Natural Science Edition), 2015, 43(3): 132-140.[曹馨月, 黃嬡媛, 趙驍, 等.蘋果蠹蛾性信息素結(jié)合蛋白Ⅱ (CpomPBP2) 基因的克隆及原核表達[J].西北農(nóng)林科技大學(xué)學(xué)報 (自然科學(xué)版), 2015, 43 (3): 132-140]

Chen L, Li HL, Zhou YX,etal.cDNA cloning, tissue expression and ligand binding characteristics of odorant-binding protein 2 from the oriental fruit fly,Bactroceradorsalis(Diptera: Tephritidae) [J].ActaEntomologicaSinica, 2013, 56 (6): 612-621.[陳玲, 李紅亮, 周宇翔, 等.桔小實蠅氣味結(jié)合蛋白BdorOBP2的cDNA克隆、組織表達及配基結(jié)合特性[J].昆蟲學(xué)報, 2013, 56 (6): 612-621]

Chen WY, Wang SS, Li DL,etal.Structure activity relationship studies on novel acylpiperidine compounds based on insect OBP1[J].ChemicalJournalofChineseUniversities, 2013, 34 (12): 2798-2805.[陳文雅, 王珊珊, 李冬玲, 等.基于昆蟲氣味結(jié)合蛋白的新型?；哙ゎ惢衔锏臉?gòu)效關(guān)系[J].高等學(xué)校化學(xué)學(xué)報, 2013, 34 (12): 2798-2805]

Damberger F, Nikonova L, Horst R,etal.NMR characterization of a pH-dependent equilibrium between two folded solution conformations of the pheromone-binding protein fromBombyxmori[J].ProteinScience, 2000, 9, 1038-1041.

Danty E, Arnold G, Huet JC.Separation characmon and sexual heterogeneity of multiple putative odorant-binding proteins in the honeybeeApismelliferaL.(Hymenoptera: Apidea) [J].ChemicalSenses, 1998, 23 (1) : 83-91

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

Forstner M, Breer H, Krieger J.A receptor and binding protein interplay in the detection of a distinct pheromone component in the silkmothAntheraeapolyphemus[J].InternationalJournalofBiologicalSciences, 2009, 5: 745-757.

Garczynski SF, Coates BS, Unruh TR,etal.Application ofCydiapomonellaexpressed sequence tags: Identification and expression of three general odorant binding proteins in codling moth [J].InsectSci., 2013, 20(5): 559-574.

Gong DP, Zhang HJ, Zhao P,etal.The odorant binding protein gene family from the genome of silkworm,Bombyxmori[J].BMCGenomics, 2009a, 10 (1): 332.

Gong ZJ, Zhou WW, Yu HZ,etal.Cloning, expression and functional analysis of a general odorant-binding protein 2 gene of the rice striped stem borer,Chilosuppressalis(Walker) (Lepidoptera: Pyralidae) [J].InsectMolecularBiology, 2009b, 18: 405-417.

Gu SH, Wang WX, Wang GR,etal.Functional characterization and immunolocalization of odorant binding protein 1 in the lucerne plant bug,Adelphocorislineolatus(Goeze) [J].ArchivesofInsectBiochemistryandPhysiology,2011, 77: 81-98.

Gu SH, Zhou JJ, Wang GR,etal.Sex pheromone recognition and immunolocalization of three pheromone binding proteins in the black cutworm mothAgrotisipsilon[J].InsectBiochemistryandMolecularBiology, 2013, 43: 237-251.

Guo H, Huang LQ, Pelosi P,etal.Three pheromone-binding proteins help segregation between two Helicoverpa species utilizing the same pheromone components [J].InsectBiochemistryandMolecularBiology, 2012, 42: 708-16.

He X, Tzotzos G, Woodcock C,etal.Binding of the general odorant binding protein ofBombyxmoriBmorGOBP2 to the moth sex pheromone components [J].JournalofChemicalEcology, 2010, 36: 1293-1305.

Hua JF, Zhang S, Cui JJ,etal.Identification and binding characterization of three odorant binding proteins and one chemosensory protein fromApolyguslucorum(Meyer-Dur) [J].JournalofChemicalEcology, 2012, 38: 1163-1170.

Ji P, Liu JT, Gu SH,etal.Expression and binding specificity analysis of odorant binding protein AlucOBP7 fromApolyguslucorum(Hemiptera: Miridae) [J].ActaEntomologicaSinica, 2013, 56 (6): 575-583.[紀萍, 劉靖濤, 谷少華, 等.綠盲蝽氣味結(jié)合蛋白AlucOBP7的表達及氣味結(jié)合特性[J].昆蟲學(xué)報, 2013, 56 (6): 575-583]

Ji T, Shen F, Liang Q,etal.Cloning, prokaryotic expression and tissue expression profiling of an OBP3 gene in the Chinese honeybee,Apisceranacerana(Hymenoptera: Apidae) [J].ActaEntomologicaSinica, 2014, 57 (8): 897-904.[吉挺, 沈芳, 梁勤, 等.中華蜜蜂OBP3基因的克隆、原核表達及組織表達譜[J].昆蟲學(xué)報, 2014, 57 (8): 897-904]

Ju Q, Li X, Jiang XJ,etal.Transcriptome and tissue log pecific expression analysis of obp and csp genes in the dark black chafer [J].ArchivesofInsectBiochemistry&Physiology, 2014, 87 (4), 177-200.

Ju Q, Qu MJ, Wang Y,etal.Molecular and biochemical characterization of two odorant-binding proteins from dark black chafer,Holotrichiaparallela[J].Genome, 2012, 55: 537-546.

Kowcun A, Honson N, Plettner E.Olfaction in the gypsy moth,Lymantriadispareffect of pH, ionic strength, and reductants on pheromone transport by pheromone-binding proteins [J].JournalofBiologicalChemistry, 2001, 276: 44770-44776.

Krieger J, Breer H.Olfactory reception in invertebrates [J].Science, 1999, 286: 720-728.

Krieger J, Breer H.Transduction mechanisms of olfactory sensory neurons [J].InsectPheromoneBiochemistryandMolecularBiology, 2003, 593-607.

Krieger J, von Nickisch-Rosenegk E, Mameli M,etal.Binding proteins from the antennae ofBombyxmori[J].InsectBiochemistryandMolecularBiology, 1996, 26: 297-307.

Kruse SW, Zhao R, Smith DP,etal.Structure of a specific alcohol-binding site defined by the odorant binding protein LUSH fromDrosophilamelanogaster [J].NatureStructural&MolecularBiology,2003, 10: 694-700.

Lartigue A, Gruez A, Briand L,etal.Sulfur single-wavelength anomalous diffraction crystal structure of a pheromone-binding protein from the honeybeeApismelliferaL [J].TheJournalofBiologicalChemistry, 2004, 279: 4459-64.

Lartigue A, Gruez A, Spinelli S,etal.The crystal structure of a cockroach pheromone-binding protein suggests a new ligand binding and release mechnism [J].TheJournalofBiologicalChemistry,2003, 278: 30213-30218.

Laughlin JD, Ha TS, Jones DNM,etal.Activation of pheromone-sensitive neurons is mediated by conformational activation of pheromone-binding protein [J].Cell, 2008, 133: 1255-1265.

Leal WS, Chen AM, Ishida Y,etal.Kinetics and molecular properties of pheromone binding and release [J].ProceedingsoftheNationalAcademyofSciencesoftheUnitedStatesofAmerica, 2005b, 102: 5386-5391.

Leal WS.Odorant reception in insects: Roles of receptors, binding proteins, and degrading enzymes [J].AnnualReviewofEntomology, 2013, 58: 373-91.

Leal WS.Pheromone reception [J].TopicsinCurrentChemistry, 2005a, 240: 1-36.

Lescop E, Briand L, Pernollet JC,etal.Structural basis of the broad specificity of a general odorant-binding protein from the honeybee [J].Biochemistry, 2009, 48: 2431-2441.

Li DZ, Yu GQ, Yi SC,etal.Structure-based analysis of the ligand-binding mechanism for DhelOBP21, a C-minus odorant binding protein, fromDastarcushelophoroides(Fairmaire; Coleoptera: Bothrideridae) [J].InternationalJournalofBiologicalSciences, 2015, 11 (11): 1281-1295.

Li F, Prestwieh GD.Expression and characterization of a Lepidopteran general odorant-binding protein [J].InsectBiochemistryandMolecularBiology, 1997, 27: 405-412.

Li HL, Zhang LY, Zhuang SL,etal.Interpretation of odorant binding function and mode of general odorant binding protein ASP2 in Chinese Honeybee (Apisceranacerana) [J].ScientiaAgriculturaSinica, 2013, 46 (1): 154-161.[李紅亮, 張林雅, 莊樹林, 等.中華蜜蜂普通氣味結(jié)合蛋白ASP2 的氣味結(jié)合功能模式分析 [J].中國農(nóng)業(yè)科學(xué), 2013, 46 (1): 154-161]

Li S, Picimbon JF, Ji S,etal.Multiple functions of an odorant-binding protein in the mosquitoAedesaegypti[J].BiochemicalandBiophysicalResearchCommunications, 2008, 372: 464-468.

Li WH, Huang XL, Wang D,etal.Molecular cloning and immunofluorescence localization of sex pheromone binding protein 2 fromAtrijuglanshetaohei(Lepidoptera: Heliodinidae) [J].ActaEntomologicaSinica, 2015, 58 (10) : 1054-1062.[李文海, 黃興龍, 王敦, 等.核桃舉肢蛾性信息素結(jié)合蛋白2的分子克隆和免疫熒光定位[J].昆蟲學(xué)報, 2015, 58 (10) : 1054-1062]

Liu NY, Liu CC, Dong SL.Functional differentiation of pheromone-binding proteins in the common cutwormSpodopteralitura[J].Comp.Biochem.Phys.A, 2013, 165: 254-262.

Liu Y, Gu S, Zhang Y,etal.Candidate olfaction genes identified within theHelicoverpaarmigeraantennal transcriptome [J].PLoSONE, 2012, 7: e48260.

Mao Y, Xu X, Xu W,etal.Crystal and solution structures of an odorant-binding protein from the southern house mosquito complexed with an oviposition pheromone [J].ProceedingsoftheNationalAcademyofSciences,USA, 2010, 107: 19102-19107.

Matsuo T, Sugaya S, Yasukawa J,etal.Odorant-binding proteins OBP57d and OBP57e affect taste perception and host-plant preference inDrosophilasechellia[J].PLoSBiology, 2007, 5 (5): e118.

Mohanty S, Zubkov S, Gronenborn M.The solution NMR structure ofAntheraeapolyphemusPBP provides new insight into pheromone recognition by pheromone-binding proteins [J].JournalofMolecularBiology, 2004, 337: 443-451.

Mombaerts P.Seven-transmembrane proteins as odorant and chemosensory receptors [J].Science, 1999, 286 (5440): 707-711.

Pelosi P, Zhou J, Ban L,etal.Soluble proteins in insect chemical communication [J].CellularandMolecularLifeSciences, 2006, 63: 1658-1676.

Qin Z, Ran YH, Zhi ZJ,etal.Cloning and expression analysis of an odorant binding protein geneAsinOBP1 fromAnophelessinensis(Diptera: Culicidae) [J].ActaEntomologicaSinica, 2014, 57 (11): 1289-1298.[秦贈, 冉永紅, 支中婧, 等.中華按蚊氣味結(jié)合蛋白基因AsinOBP1的克隆和表達分析[J].昆蟲學(xué)報, 2014, 57 (11): 1289-1298]

Ren ZZ, Liu XL, Hu MY.Structure and function of insect Olfactory proteins [J].ChineseJournalofBiochemistryandMolecularBiology, 2010, 26 (6): 531-537.[任珍珍, 柳曉磊, 胡美英.昆蟲嗅覺相關(guān)蛋白的結(jié)構(gòu)和功能[J].中國生物化學(xué)與分子生物學(xué)報, 2010, 26 (6): 531-537]

Sandler BH, Nikonova L, Leal WS,etal.Sexual attraction in the silkworm moth: Structure of the pheromone-binding-protein-bombykol complex [J].ChemistryandBiology, 2000, 7: 143-151.

Schneider D.Insect olfaction: deciphering system for chemical messages [J].Science, 1969, 163 (3871): 1031-1037.

Schultze A, Schymura D, Forstner M.Expression pattern of a“Plus-C”class odorant binding protein in the antenna of the malaria vectorAnophelesgambiae[J].InsectMol.Biol., 2012, 21 (2) : 187-195.

Song YQ, Xie XC, Dong JF,etal.cDNA cloning, expression profiling and binding properties of odorantbinding protein GmolOBP3 in the oriental fruit moth,Grapholitamolesta( Lepidoptara: Tortricidae) [J].ActaEntomologicaSinica, 2014, 57 (3): 274-285.[宋月芹, 解幸承, 董鈞鋒, 等.梨小食心蟲氣味結(jié)合蛋白GmolOBP3的cDNA克隆、表達譜及結(jié)合特性分析[J].昆蟲學(xué)報, 2014, 57 (3): 274-285]

Spinelli S, Lagarde A, Iovinella I,etal.Crystal structure ofApismelliferaOBP14, a C-minus odorant-binding protein, and its complexes with odorant molecules [J].InsectBiochemistryandMolecularBiology, 2012, 42 (1): 41-50.

Sun H, Shu M, Zhao LX,etal.Homology modeling of pheromone binding protein ofPectinophoragossypiella[J].JournalofEnvironmentalEntomology, 2013, 35 (6): 749-753.[孫浩, 舒茂, 趙雷修, 等.棉紅鈴蟲信息素結(jié)合蛋白的同源模建[J].環(huán)境昆蟲學(xué)報, 2013, 35 (6) : 749-753]

Sun HY, Yin J, Feng HL,etal.Expression, purification and binding characteristics of general odorant binding proteinⅠ (GOBP1) from themeadow moth,Loxostegesticticalia(Linnaeus) (Lepidoptera: Pyralididae) [J].ActaEntomologicaSinica, 2011, 54 (4): 381-389.[孫紅巖, 尹姣, 馮紅林, 等.草地螟普通氣味結(jié)合蛋白Ⅰ(Lsti-GOBP1)蛋白表達純化及結(jié)合特性分析[J].昆蟲學(xué)報, 2011, 54 (4): 381 -389]

Sun MJ, Liu Y, Wang GR.Expression patterns and binding properties of three pheromone binding proteins in the diamondback moth,Plutellaxyllotella[J].JournalofInsectPhysiology, 2013, 59: 46-55.

Tegoni M, Campanacci V, Cambillau C.Structural aspects of sexual attraction and chemical communication in insects [J].TrendsinBiochemicalSciences, 2004, 29: 257-264.

Tian Z, Liu J, Zhang Y.Structural insights intoCydiapomonellapheromone binding protein 2 mediated prediction of potentially active semiochemicals [J].ScientificReports, 2016, 6: 1-16.

Vandermoten S, Francis F, Haubruge E,etal.Conserved odorant-binding proteins from aphids and eavesdropping predators [J].PLoSONE, 2011, 6: e23608.

Vogel H, Heidel AJ, Heckel DG,etal.Transcriptome analysis of the sex pheromone gland of the noctuid mothHeliothisvirescens[J].BMCGenomics.2010, 11: 29.

Vogt RG, Riddiford LM.Pheromone binding and inactivation by moth antennae [J].Nature, 1981, 293 (5828): 161-163.

Vogt RG.Molecular basis of pheromone detection in insects.In: Gilbert L, Iatro K, Gill S, ed.Comprehensive Insect Physiology, Biochemistry, Pharmacology and Molecular Biology.Vol.3 [C].London: Elsevier; 2005: 753-804.

Wang GR, Wu KM, Guo YY.Cloning, expression and immunocytochemical localization of a general odorant binding protein gene fromHelicoverpaarmigera(Hubner) [J].InsectBiochemistryandMolecularBiology, 2003, 33: 115-124.

Wei D, Ye ZF, Gao JQ,etal.Molecular cloning and functional identification of a Minus-C odorant binding protein from the rice striped stem borer,Chilosuppressalis(Lepidoptera: Pyralidae) [J].ActaEntomologicaSinica, 2013, 56 (7): 754-764.[魏丹, 葉占峰, 高建清, 等.二化螟Minus-C氣味結(jié)合蛋白的分子克隆及功能鑒定[J].昆蟲學(xué)報, 2013, 56 (7): 754-764]

Wogulis M, Morgan T, Ishida Y,etal.The crystal structure of an odorant binding protein fromAnophelesgambiae: Evidence for a common ligand release mechanism [J].BiochemicalandBiophysicalResearchCommunications, 2006, 339 (1): 157-164.

Wojtasek H, Leal WS.Conformational change in the pheromone-binding protein fromBombyxmoriinduced by pH and by interaction with membranes [J].JournalofBiologicalChemistry, 1999, 274: 30950-30956.

Xiu H, Zheng BH, Yu JZ,etal.Genome-wide identification and characterization of odorant-binding protein (obp) genes in the malaria vectorAnophelessinensis, (Diptera: Culicidae) [J].InsectScience, 2016, 23(3): 366-376.

Xu X, Xu W, Rayo J,etal.NMR structure of navel orangeworm moth pheromone-binding protein (AtraPBPl): Implications for pH-sensitive pheromone detection [J].Biochemistry, 2010, 49: 1469-1476.

Yin J, Feng H, Sun H,etal.Functional analysis of general odorant binding protein 2 from the Meadow Moth,LoxostegesticticalisL.(Lepidoptera: Pyralidae) [J].PLoSONE, 2012, 7: e33589.

Yu F, Zhang S, Zhang L,etal.Intriguing similarities between two novel odorant-binding proteins of locusts [J].BiochemicalandBiophysicalResearchCommunications, 2009, 385: 369-374.

Zhang S.Identification and Expression Profile Analysis of Odorant-binding Proteins Genes Based on Transcriptome in Grasshopper,Oedaleusasiaticus[D].Hohhot: Journal of Inner Mongolia Agricultural University, 2015: 31-32.[張碩.基于轉(zhuǎn)錄組的亞洲小車蝗氣味結(jié)合蛋白基因的鑒定及其表達譜分析[D].內(nèi)蒙古農(nóng)業(yè)大學(xué), 2015:31-32]

Zhang ZC, Wang MQ, Zhang GA.Molecular cloning and expression of pheromone binding protein 1 from the diamondback moth,Plutellaxylostella[J].EntomologiaExperimentalisetApplicata, 2009, 133: 136-145.

Zhao H, Luo Y, Lee J,etal.The odorant-binding protein gene obp11 shows different spatiotemporal roles in the olfactory system ofApismelliferaligusticaandApisceranacerana[J].Sociobiology, 2013, 60 (4): 429-435.

Zhao HX, Zeng XN, Liang Q,etal.Research on honeybee odorant-binding proteins biochemical characteristics, phyletic evolution, expression location and physiological function [J].JournalofEnvironmentalEntomology, 2015, 37 (2): 420-425.[趙紅霞, 曾鑫年, 梁勤, 等.蜜蜂氣味結(jié)合蛋白的生化特性、系統(tǒng)進化、表達部位和生理功能的研究[J].環(huán)境昆蟲學(xué)報, 2015, 37 (2): 420-425]

Zhao X, Han LL, Wang L,etal.Sequence characteristic and expression profiling of odorant binding protein 7 gene fromAcyrthosiphonsolani[J].ChineseJournalofBiologicalControl, 2014, 30 (5): 639-646.[趙雪, 韓嵐嵐, 王玲, 等.茄無網(wǎng)蚜Acyrthosiphonsolani氣味結(jié)合蛋白OBP7的序列特點及表達譜分析[J].中國生物防治學(xué)報, 2014, 30 (5): 639-646]

Zhong T, Yin J, Deng SS,etal.Fluorescence competition assay for the assessment of green leaf volatiles and trans-β-farnesene bound to three odorant-binding proteins in the wheat aphidSitobionavenae(Fabricius) [J].JournalofInsectPhysiology, 2012, 58: 771-778.

Zhou JJ, Robertson G, He XL,etal.Characterisation ofBombyxmoriodorant-binding proteins reveals that a general odorant-binding protein discriminates between sex pheromone components [J].JournalofMolecularBiology.2009, 389: 529-545.

Zhou JJ, Vieira FG, He XL,etal.Genome annotation and comparative analyses of the odorant-binding proteins and chemosensory proteins in the pea aphidAcyrthosiphonpisum[J].InsectMolecularBiology, 2010, 19 (Suppl.2): 113-122.

Zhou JJ, Zhang GA, Huang W,etal.Revisiting the odorant-binding protein LUSH ofDrosophilamelanogaster: Evidence for odour recognition and discrimination [J].FEBSLetters, 2004, 558 (1-3): 23-26.

Zhu JY, Zhang LF, Ze SZ,etal.Identification and tissue distribution of odorant binding protein genes in the beet armyworm,Spodopteraexigua[J].JournalofInsectPhysiology, 2013, http: //dx.doi.org/10.1016/jjinsphys.2013.02.011.

Recent advances in odorant binding proteins of insects

ZHANG Zhi-Ke1*，ZHANG Ye2，WU Sheng-Yong3，LEI Zhong-Ren3, 4*

(1.Ningxia Key Laboratory of Plant Diseases and Pests Control, Institute of Plant Protection, Ningxia Academy of Agriculture and Forestry Sciences, Yinchuan 750002, China; 2.Institute of Plant Protection, Shanxi Academy of Agricultural Sciences, Taiyuan 030031, China; 3.State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China; 4.Fujian-Taiwan Joint Centre for Ecological Control of Crop Pests, Fuzhou 350002, China)

Olfactory plays important roles in the behavior of insects, illustrating the olfactory mechanism of insects will be useful to regulate insect behaviors and develop new strategy for pest control.Odorant binding proteins is one class important proteins in insect olfactory system.In recent years, with the rapid development of genome and transcriptome, more and more odorant binding proteins were identified, among them, some proteins’ functions were proved gradually, which are beneficial to reveal the machenism of insect olfactory.This paper summarized the classes, structure, expression, distribution，three-dimensional model, physiological functions of odorant binding proteins, which could provide reference for identifying more odorant binding proteins and studying their functions, lay the foundation for revealling furtherly chemical communication mechanism between insect and environment.

Insect olfactory; odorant binding protein; structural characteristics; expression and distribution; physiological function

張治科，張燁，吳圣勇,等.昆蟲氣味結(jié)合蛋白研究進展[J].環(huán)境昆蟲學(xué)報，2017,39(3):713-720.

國家自然科學(xué)基金(31660621)；寧夏農(nóng)林科學(xué)院科技創(chuàng)新先導(dǎo)資金(NKYJ-17-05，NKYJ-15-15)；寧夏自然科學(xué)基金(NZ15128)；自治區(qū)重點研發(fā)計劃重大項目(2016BZ09)；一二三產(chǎn)業(yè)融合發(fā)展科技創(chuàng)新示范項目(YES-16-03)；國家重點研發(fā)計劃項目(2016YEC1201200)

張治科，男，1980年生，博士，副研究員，主要從事昆蟲生態(tài)與綜合防治以及昆蟲化學(xué)生態(tài)學(xué)研究，E-mail: zhangzhike98@163.com

*通訊作者Author for correspondence, E-mail: zhangzhike98@163.com; leizhr@sina.com

Received: 2016-04-14; 接受日期Accepted: 2016-09-18

Q965;S433

A

1674-0858(2017)03-0713-08