Functional characterization of a geranylgeranyl diphosphate synthase in the leaf beetle Monolepta hieroglyphica

• Published in: Archives of insect biochemistry and physiology Vol. 115; no. 2; pp. e22088 - n/a
• Main Authors: Song, Xuan, Liu, Chang, Dhiloo, Khalid H., Yi, Chao‐qun, Zhang, Tian‐tao, Zhang, Yong‐jun
• Format: Journal Article
• Language: English
• Published: United States Wiley Subscription Services, Inc 01.02.2024
• Subjects: Beetles; Binding sites; Biosynthesis; Bonding strength; catalytic activity; Chrysomelidae; dimethylallyltranstransferase; Functional plasticity; geranylgeranyl diphosphate synthase; Hydrogen bonding; Hydrogen bonds; insect biochemistry; Insects; isoprenoids; kinetics; Leaves; Mevalonate pathway; Molecular docking; Monolepta hieroglyphica; Phylogeny; Prenyltransferases; recombinant expression; Substrates; Monolepta hieroglyphica; recombinant expression; molecular docking; geranylgeranyl diphosphate synthase; catalytic activity
• ISSN: 0739-4462; 1520-6327; 1520-6327
• DOI: 10.1002/arch.22088

Geranylgeranyl diphosphate synthase (GGPPS) as the short‐chain prenyltransferases for catalyzing the formation of the acyclic precursor (E)‐GGPP has been extensively investigated in mammals, plants, and microbes, but its functional plasticity is poorly understood in insect species. Here, a single GGPPS in leaf beetle Monolepta hieroglyphica, MhieGGPPS, was functionally investigated. Phylogenetic analysis showed that MhieGGPPS was clustered in one clade with homologs and had six conserved motifs. Molecular docking results indicated that binding sites of dimethylallyl diphosphate (DMAPP), (E)‐geranyl pyrophosphate (GPP), and (E)‐farnesyl pyrophosphate (FPP) were in the chain‐length determination region of MhieGGPPS, respectively. In vitro, recombiant MhieGGPPS could catalyze the formation of (E)‐geranylgeraniol against different combinations of substrates including isopentenyl pyrophosphate (IPP)/DMAPP, IPP/(E)‐GPP, and IPP/(E)‐FPP, suggesting that MhieGGPPS could not only use (E)‐FPP but also (E)‐GPP and DMAPP as the allylic cosubstrates. In kinetic analysis, the (E)‐FPP was most tightly bound to MhieGGPPS than that of others. It was proposed that MhieGGPPS as a multifunctional enzyme is differentiated from the other GGPPSs in the animals and plants, which only accepted (E)‐FPP as the allylic cosubstrate. These findings provide valuable insights into understanding the functional plasticity of GGPPS in M. hieroglyphica and the novel biosynthesis mechanism in the isoprenoid pathway. A geranylgeranyl diphosphate synthase, MhieGGPPS, was identified in the leaf beetle. Substrates strongly bound to MhieGGPPS by the formation of hydrogen bonds with the key residues in the CLD. MhieGGPPS was multifunctional in catalytic activities. Highlights A geranylgeranyl diphosphate synthase, MhieGGPPS, was identified in the leaf beetle. Substrates strongly bound to MhieGGPPS by the formation of hydrogen bonds with the key residues in the CLD. MhieGGPPS was multifunctional in catalytic activities.