A Highly Expressed Antennae Odorant-Binding Protein Involved in Recognition of Herbivore-Induced Plant Volatiles in Dastarcus helophoroides
Abstract
:1. Introduction
2. Results
2.1. Analyses of HIPVs of P. massoniana Lamb
2.2. Odorant-Binding Proteins in D. helophoroides
2.3. Spatio-Temporal Expression Patterns of DhelOBPs
2.4. AlphaFold2-Based Modeling and Molecular Docking of DhelOBPs
2.5. Binding Characteristic of Recombinant DhelOBPs
2.6. Behavioral Response of D. helophoroides to HIPVs with the Best Binding Affinities
2.7. Binding Conformation of DhelOBP4 with HIPVs
3. Discussion
4. Methods and Materials
4.1. Insect Culturing and Pine Woods Collection
4.2. Collection and Analysis of P. massoniana HIPVs
4.3. Classification of DhelOBPs
4.4. Tissue Sampling, RNA Extraction, and cDNA Synthesis
4.5. Spatio-Temporal Expression Analyses
4.6. AlphaFold2-Based Modeling and Molecular Docking
4.7. Subcloning, Expression, and Purification of Target DhelOBPs
4.8. Fluorescence Competitive Binding Assays
4.9. Y-Tube Olfactometer Experiments
4.10. Synthesis and Microinjection of dsRNA
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Number | Substance | Retention Time (min) | PubChem CID | CAS Number | 2-D Structures |
---|---|---|---|---|---|
1 | α-Fenchene | 9.260 | 28930 | 471-84-1 | |
2 | p-Cymene | 11.454 | 7463 | 99-87-6 | |
3 | γ-Terpinene | 12.389 | 7461 | 99-85-4 | |
4 | Terpinolene | 13.178 | 11463 | 586-62-9 | |
5 | Fenchone | 13.312 | 14525 | 7787-20-4 | |
6 | Camphor | 14.939 | 2537 | 76-22-2 | |
7 | Pinocamphone | 15.272 | 6427105 | 547-60-4 | |
8 | Isopinocamphone | 15.707 | 84532 | 14575-93-0 | |
9 | Terpinen-4-ol | 15.820 | 11230 | 562-74-3 | |
10 | α-Terpineol | 16.186 | 17100 | 98-55-5 | |
11 | (-)-Verbenone | 16.519 | 92874 | 1196-01-6 |
Ligands | DhelOBP4 | DhelOBP5 | DhelOBP6 | DhelOBP14 | DhelOBP18 | DhelOBP20 |
---|---|---|---|---|---|---|
α-Fenchene | −5.9 | −5.4 | −6 | −6.4 | −6.3 | −6.5 |
p-Cymene | −6.7 | −6.2 | −6.1 | −7.3 | −6.8 | −6.5 |
γ-Terpinene | −6.3 | −6.1 | −6.3 | −7.2 | −6.7 | −6.6 |
Terpinolene | −6.5 | −6.2 | −6.2 | −7.4 | −6.9 | −6.4 |
Fenchone | −5.9 | −6.2 | −6.6 | −6.7 | −6.7 | −6.5 |
Camphor | −5.6 | −5.5 | −6.3 | −6.3 | −6.1 | −6.8 |
Pinocamphone | −6.1 | −6.2 | −6.8 | −7.0 | −6.5 | −6.8 |
Isopinocamphone | −6.2 | −5.7 | −6.6 | −6.7 | −6.5 | −6.6 |
Terpinen-4-ol | −6.2 | −6.1 | −6.2 | −7.1 | −6.9 | −6.3 |
α-Terpineol | −6 | −6.3 | −6.3 | −7.1 | −6.7 | −6.3 |
(-)-Verbenone | −6.2 | −6 | −6.5 | −6.4 | −6.4 | −6.7 |
Ligands | DhelOBP4 | DhelOBP5 | DhelOBP6 | DhelOBP14 | DhelOBP18 | DhelOBP20 | ||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
pH 7.4 | pH 5.0 | pH 7.4 | ||||||||||||
IC50 | Ki | IC50 | Ki | IC50 | Ki | IC50 | Ki | IC50 | Ki | IC50 | Ki | IC50 | Ki | |
p-Cymene | 13.0 ± 0.9 | 12.0 ± 0.9 | >50 | - | >50 | - | ud | ud | >50 | - | >50 | - | >50 | - |
γ-Terpinene | 11.0 ± 0.3 | 10.1 ± 0.3 | >50 | - | >50 | - | ud | ud | >50 | - | >50 | - | >50 | - |
Terpinolene | 15.7 ± 1.8 | 14.5 ± 1.7 | >50 | - | >50 | - | ud | ud | >50 | - | >50 | - | >50 | - |
Fenchone | 23.8 ± 5.1 | 22.0 ± 4.7 | 47.4 ± 2.6 | 43.8 ± 2.4 | >50 | - | ud | ud | >50 | - | >50 | - | >50 | - |
Camphor | 18.0 ± 2.4 | 16.6 ± 2.2 | >50 | - | >50 | - | ud | ud | >50 | - | >50 | - | >50 | - |
Terpinen-4-ol | 27.8 ± 3.5 | 25.7 ± 3.2 | >50 | - | >50 | - | ud | ud | >50 | - | >50 | - | >50 | - |
α-Terpineol | 10.4 ± 0.1 | 9.6 ± 0.1 | >50 | - | >50 | - | ud | ud | >50 | - | >50 | - | >50 | - |
(-)-Verbenone | 23.1 ± 3.9 | 21.3 ± 3.6 | >50 | - | >50 | - | ud | ud | >50 | - | >50 | - | >50 | - |
Ligands | Hydrophilic Residues | Hydrophobic Residues |
---|---|---|
α-Fenchene | - | Leu 11, Leu 32, Met 46, Ile 47, Met 50, Phe 126 |
p-Cymene | Tyr 90 | Val 7, Phe 54, Val 56, Ala 68, Phe 71, Leu 72, Met 86 |
γ-Terpinene | Tyr 90 | Val 7, Phe 54, Val 56, Ala 68, Phe 71, Leu 72, Met 86 |
Terpinolene | His 51 | Val 7, Leu 11, Met 50, Phe 54, Phe 71, Phe 126 |
Fenchone | Tyr 90 | Val 56, Ala 68, Phe 71, Leu 72, Met 86, Ile 89 |
Camphor | His 51, Gly 113 | Ile 89, Leu 109, Ala 110, Ile 114, Pro 125, Phe 126 |
Pinocamphone | - | Leu 11, Leu 32, Ile 47, Met 50, Phe 54, Phe 126 |
Isopinocamphone | - | Leu 11, Leu 32, Met 46, Ile 47, Met 50, Phe 126 |
Terpinen-4-ol | Tyr 90 | Phe 54, Val 56, Ala 68, Phe 71, Leu 72, Met 86 |
α-Terpineol | His 51, Tyr 90, Cys 93 | Val 56, Ile 89, Ala 94, Leu 109, Ala 110, Phe 126 |
(-)-Verbenone | - | Leu 11, Leu 32, Met 46, Ile 47, Met 50, Phe 126 |
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Yi, S.-C.; Wu, Y.-H.; Yang, R.-N.; Li, D.-Z.; Abdelnabby, H.; Wang, M.-Q. A Highly Expressed Antennae Odorant-Binding Protein Involved in Recognition of Herbivore-Induced Plant Volatiles in Dastarcus helophoroides. Int. J. Mol. Sci. 2023, 24, 3464. https://doi.org/10.3390/ijms24043464
Yi S-C, Wu Y-H, Yang R-N, Li D-Z, Abdelnabby H, Wang M-Q. A Highly Expressed Antennae Odorant-Binding Protein Involved in Recognition of Herbivore-Induced Plant Volatiles in Dastarcus helophoroides. International Journal of Molecular Sciences. 2023; 24(4):3464. https://doi.org/10.3390/ijms24043464
Chicago/Turabian StyleYi, Shan-Cheng, Yu-Hang Wu, Rui-Nan Yang, Dong-Zhen Li, Hazem Abdelnabby, and Man-Qun Wang. 2023. "A Highly Expressed Antennae Odorant-Binding Protein Involved in Recognition of Herbivore-Induced Plant Volatiles in Dastarcus helophoroides" International Journal of Molecular Sciences 24, no. 4: 3464. https://doi.org/10.3390/ijms24043464