Frontiers in Physiology
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2025
·
IF 3.4
Long Liu,Yangdi Li,Hua Yang,Fan Wang,Qiong Huang
Odorant-binding proteins (OBPs) are important for insects to discriminate, bind and transport odorants, such as pheromones and host plant volatiles. Herein, the Minus-C OBP (CbuqOBP1) was characterized from Cyrtotrachelus buqueti , one of the most important pests in bamboo plantations. CbuqOBP1 showed significantly higher transcription levels in the adult stage and was most highly expressed in the head of both sexes, the thorax and antenna of the male, indicating that it plays important roles in chemosensory behavior of adults and may also function in other biological processes. Fluorescence competitive binding assays showed that CbuqOBP1 displayed broad binding capabilities and strong affinities to phenol ( K i = 10.49 μM) and benzothiazole ( K i = 11.11 μM) among 8 C. buqueti volatiles. CbuqOBP1 also showed high binding affinity to the main volatile of the host plant Neosinocalamus affinis (linalool, K i = 13.41 μM). The docking results indicated that hydrophobic interactions were the prevailing forces between CbuqOBP1 with these three ligands. Additionally, several amino acid residues were significantly overlapped and contributed to the interactions with the ligands. The combined results suggest that CbuqOBP1 may play dual roles in binding volatile compounds from the host plant and the same species and will be helpful to developing new pest-control strategies.
研究领域:
昆虫学
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分子生物学
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DOI
PubMed
高影响因子
Insects
·
2024
·Vol. 15
·期 2
·111
·
IF 2.7
Hua Yang,Long Liu,Fan Wang,Wei Yang,Qiong Huang,Nanxi Wang,Hongling Hu
Simple SummaryThe bamboo weevil beetle,Cyrtotrachelus buqueti(Coleoptera: Curculionidae) is the primary bamboo pest and seriously influences the development of the bamboo industry. Sensory neuron membrane proteins (SNMPs) play important roles in insect pheromone communication. However, SNMPs ofC. buquetiare still uncharacterized. With the aim of developing control techniques to control this pest, one novelSNMPfromC. buqueti,CbuqSNMP1b, was functionally characterized in this study. The results indicated thatCbuqSNMP1bwas predominantly expressed in the antennae of both sexes and showed significantly higher transcription levels in the adult stage, suggesting thatCbuqSNMP1bis involved in the process of olfaction. Fluorescence binding assays revealed that CbuqSNMP1b could bind to three out of fourteen volatile compounds emitted fromC. buquetiand showed the strongest binding affinity to dibutyl phthalate, demonstrating the role of SNMP1 in detecting odorants. Molecular docking was performed to further understand the binding mode between CbuqSNMP1b and these three ligands, and the results showed that hydrophobic interactions were the prevailing forces within the binding cavities of CbuqSNMP1b. The results of this study will be helpful to understand the function of SNMP1 inC. buquetiand lay a foundation for developing new methods to control this pest.AbstractSensory neuron membrane proteins (SNMPs) play important roles in insect chemoreception and SNMP1s have been reported to be essential in detecting sex pheromones inDrosophilaand some lepidopteran species. However, SNMPs forCyrtotrachelus buqueti(Coleoptera: Curculionidae), a major insect pest of bamboo plantations, remain uncharacterized. In this study, a novelSNMPgene,CbuqSNMP1b, fromC. buquetiwas functionally characterized. The expression ofCbuqSNMP1bwas significantly higher in antennae than in other tissues of both sexes and the expression level was significantly male-biased. Additionally,CbuqSNMP1bshowed significantly higher transcription levels in the adult stage and very low transcription levels in other stages, suggesting thatCbuqSNMP1bis involved in the process of olfaction. Fluorescence binding assays indicated that CbuqSNMP1b displayed the strongest binding affinity to dibutyl phthalate (Ki= 9.03 μM) followed by benzothiazole (Ki= 11.59 μM) and phenol (Ki= 20.95 μM) among fourteenC. buquetivolatiles. Furthermore, molecular docking revealed key residues in CbuqSNMP1b that interact with dibutyl phthalate, benzothiazole, and phenol. In conclusion, these findings will lay a foundation to further understand the olfactory mechanisms ofC. buquetiand promote the development of novel methods for controlling this pest.Keywords:Cyrtotrachelus buqueti;sensory neuron membrane protein;expression pattern;binding assay;molecular docking
研究领域:
昆虫学
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分子生物学
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害虫管理
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PubMed
高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2023
·Vol. 24
·期 23
·16925
·
IF 5.6
Long Liu,Fan Wang,Wei Yang,Hua Yang,Qiong Huang,Chunlin Yang,Wenkai Hui
Pheromone-binding proteins (PBPs) play important roles in binding and transporting sex pheromones. However, thePBPgenes identified in coleopteran insects and their information sensing mechanism are largely unknown.Cyrtotrachelus buqueti(Coleoptera: Curculionidae) is a major insect pest of bamboo plantations. In this study, a novelPBPgene,CbuqPBP2, fromC. buquetiwas functionally characterized.CbuqPBP2was more abundantly expressed in the antennae of both sexes than other body parts, and its expression level was significantly male-biased. Fluorescence competitive binding assays showed that CbuqPBP2 exhibited the strongest binding affinity to dibutyl phthalate (Ki= 6.32 μM), followed by styrene (Ki= 11.37 μM), among twelveC. buquetivolatiles. CbuqPBP2, on the other hand, showed high binding affinity to linalool (Ki= 10.55), the main volatile of host plantNeosinocalamus affinis. Furthermore, molecular docking also demonstrated the strong binding ability of CbuqPBP2 to dibutyl phthalate, styrene, and linalool, with binding energy values of −5.7, −6.6, and −6.0 kcal/mol, respectively, and hydrophobic interactions were the prevailing forces. The knockdown ofCbuqPBP2expression via RNA interference significantly reduced the electroantennography (EAG) responses of male adults to dibutyl phthalate and styrene. In conclusion, these results will be conducive to understanding the olfactory mechanisms ofC. buquetiand promoting the development of novel strategies for controlling this insect pest.Keywords:Cyrtotrachelus buqueti;pheromone-binding proteins;binding assay;molecular docking;RNA interference;electroantennography
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DOI
PubMed
高影响因子
PeerJ
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2019
·
IF 2.353
Hua Yang,Yan-Lin Liu,Yuan-Yuan Tao,Wei Yang,Chun-Ping Yang,Jing Zhang,Li-Zhi Qian,Hao Liu,Zhi-Yong Wang
The bamboo snout beetle Cyrtotrachelus buqueti is a widely distributed wood-boring pest found in China, and its larvae cause significant economic losses because this beetle targets a wide range of host plants. A potential pest management measure of this beetle involves regulating olfactory chemoreceptors. In the process of olfactory recognition, pheromone-binding proteins (PBPs) play an important role. Homology modeling and molecular docking were conducted in this study for the interaction between CbuqPBP1 and dibutyl phthalate to better understand the relationship between PBP structures and their ligands. Site-directed mutagenesis and binding experiments were combined to identify the binding sites of CbuqPBP1 and to explore its ligand-binding mechanism. The 3D structural model of CbuqPBP1 has six a-helices. Five of these a-helices adopt an antiparallel arrangement to form an internal ligand-binding pocket. When docking dibutyl phthalate within the active site of CbuqPBP1, a CH- π interaction between the benzene ring of dibutyl phthalate and Phe69 was observed, and a weak hydrogen bond formed between the ester carbonyl oxygen and His53. Thus, Phe69 and His53 are predicted to be important residues of CbuqPBP1 involved in ligand recognition. Site-directed mutagenesis and fluorescence assays with a His53Ala CbuqPBP1 mutant showed no affinity toward ligands. Mutation of Phe69 only affected binding of CbuqPBP1 to cedar camphor. Thus, His53 (Between α2 and α3) of CbuqPBP1 appears to be a key binding site residue, and Phe69 (Located at α3) is a very important binding site for particular ligand interactions.
研究领域:
生物信息学
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生化分析
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昆虫学
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Hua Yang,Long Liu,Ling Chen,Chunlin Yang,Qiong Huang,Nanxi Wang,Hongling Hu
Sensory neuron membrane proteins (SNMPs) play critical roles in insect olfactory system. However, functional studies outside of Drosophila remain limited, especially in Coleoptera species. In our previous study, a SNMP1 (CbuqSNMP1b) was identified from Cyrtotrachelus buqueti (Coleoptera: Curculionidae), an insect pest that seriously influence the development of the bamboo industry. Here in a membrane protein yeast two-hybrid system, protein interactions between CbuqSNMP1b as a bait protein and a cDNA library of antenna of male C. buqueti adults as prey protein were assessed. Of 29 proteins identified as putative interactors, the Minus-C odorant-binding protein (CbuqOBP1) was selected for further analysis. The interaction between CbuqSNMP1b and CbuqOBP1 was further confirmed by both the in vivo yeast spotting analysis and the in vitro glutathione-S-transferase pull-down assay. Fluorescence binding assays indicated that the interaction between CbuqSNMP1b and CbuqOBP1 could enhance the binding abilities of CbuqOBP1 to four adult C. buqueti biologically active volatiles. The knockdown of CbuqSNMP1b + CbuqOBP1 expression by RNA interference significantly reduced the behavior responses of male adults to ethyl hexanoate and trans , trans -2,4-Nonadienal. These results increase our understanding of insect SNMP1 and will aid in exploring the underlying mechanisms of CbuqSNMP1b functions in the future.
研究领域:
昆虫学
·
分子生物学
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产品关联
DOI
PubMed
高影响因子
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