Novel proteinase inhibitor from the hemolymph of soybean pest Anticarsia gemmatalis (lepidóptera: Noctuidae): Structural modeling and enzymatic kinetic

• Published in: Archives of insect biochemistry and physiology 2022-03, Vol.109 (3), p.e21864-n/a
• Main Authors: Rios‐Díez, Juan D., Meriño‐Cabrera, Yaremis, Silva‐Junior, Neilier Rodrigues, Almeida Barros, Rafael, Aguilar de Oliveira, João, Josué de Oliveira Ramos, Humberto, Goreti de Almeida Oliveira, Maria
• Format: Article
• Language: English
• Subjects: Affinity; Animals; Anticarsia gemmatalis; caterpillar; Cattle; competitive inhibition; docking; Enzymes; Glycine max; Hemolymph; Insecticide resistance; Insecticides; Larva; Lepidoptera; Modelling; Molecular docking; Molecular Docking Simulation; Molecular modelling; Moths; Natural enemies; Pest control; Pesticide resistance; Pests; Protease Inhibitors; Proteinase; Proteinase inhibitors; SERPIN; Soybeans; Toxicity; Trypsin
• ISSN: 0739-4462; 1520-6327
• DOI: 10.1002/arch.21864

New approaches are needed to reduce risks to the environment and natural enemies and to avoid or delay the onset of insecticide resistance. The use of insecticides based on proteinase inhibitors of hemolymph is an alternative for the control of Lepidoptera pests primarily by having low toxicity and short persistence in the environment. Thus, in this study, we describe the purification process and identification of protease inhibitors from hemolymph Anticarsia gemmatalis and their activities against trypsin enzymes. Furthermore, the three‐dimensional (3D) structure of the inhibitor and binding mode to trypsin enzymes was determined, and the stability of the inhibitory activity in several pHs and temperature values was evaluated. The inhibitor was characterized as a serpin family inhibitor and named A. gemmatalis hemolymph serpin inhibitor (AHSI), with an approximate mass of 38 ± 2 kDa, highly stable to temperature and pH variations, and with inhibitory capacity on bovine trypsin and gut trypsin of A. gemmatalis demonstrated by calculated Ki values and affinity energy through molecular docking, being a reversible competitive inhibitor that binds to the active site of trypsin‐like enzymes. We conclude that the AHSI inhibitor identified from the hemolymph of the soybean pest A. gemmatalis preserves the original structure of the serpin family with a good overall stereochemical quality confirmed from molecular modeling. The docking analysis showed that the reactive site of the inhibitor is in contact with the catalytic cavity of the trypsin with high‐affinity energy. The three‐dimensional structure (3D) modeling of Anticarsia gemmatalis hemolymph serpin inhibitor (AHSI) conserves the original structure of the serpin family, presenting 21% of α‐helix, 34.4% of β‐strands, and 44.6% of coiled coils. AHSI is a competitive and reversible inhibitor on the A. gemmtalis trypsin enzymes. AHSI has a higher affinity for gut trypsins of A. gemmatalis, with an approximate mass of 38 ± 2 kDa, highly stable to temperature and pH variations.