Proteomic dentification and functional characterization of venom protein HhUp140 of Habrobracon hebetor as a paralytic protein

Abstract BACKGROUND Parasitoid wasps play a crucial role in biological control of insect pests. Habrobracon hebetor (Say) is a prominent parasitoid wasp that targets a diverse array of field and storage pests, including Plodia interpunctella (Hübner). Its venom has the potential to serve as an effective pest control agent by disrupting the movement, metabolism, and development of hosts. RESULTS iTRAQ analysis of the hemolymph of P. interpunctella after parasitism by H. hebetor identified a total of 821 host response proteins. These host proteins showed significant differences in those related to neural signal transmission. Additionally, 12 potential venom proteins of H. hebetor were identified, including two homologous proteins (HhPp1 and HhUp140). Protein sequences analysis revealed HhPp1 has a total of 106 amino acids and was classified as a basic protein with a molecular mass of 11.09 kDa. HhUp140 comprises a total of 99 amino acids and was classified as an acidic protein with a molecular mass of 10.37 kDa. Both proteins were recombinantly expressed using the pET‐28a(+) vector and subsequently injected into P. interpunctella larvae. After injection the host larvae exhibited whole‐body paralysis, a diminished stress response, and even death of some individuals when the recombinant HhUp140‐His was injected. And paralysis affected the pupation process of the larvae. However, no reactions were observed following the injection of recombinant HhPp1‐His. CONCLUSION These results suggest that HhUp140 may function as a potential paralytic venom protein, while HhPp1 does not show any paralytic activity. © 2025 Society of Chemical Industry.