Characterization and in vitro expression of arginine kinase gene in the invasive western flower thrips, Frankliniella occidentalis

Comp Biochem Physiol B Biochem Mol Biol. 2019 Mar:229:51-57. doi: 10.1016/j.cbpb.2019.01.003. Epub 2019 Jan 11.

Abstract

Arginine kinase (AK) plays a critical role in insect energy metabolism and has been proposed to be a potential insecticide target for commercial exploitation. In this study, the full length cDNA encoding a typical group 1 insect AK (FoAK) was isolated from the western flower thrips (WFT), Frankliniella occidentalis (Pergande). Sequence analysis showed that FoAK contains an open reading frame of 1068 nucleotides, which encods a protein of 355 amino acid residues including the signature sequence pattern of ATP-guanidino kinases. Genomic structure analysis showed that the coding region of FoAK contains five exons connected by four introns. RT-qPCR analysis revealed that the mRNA expression of FoAK was developmentally regulated with the lowest level in prepupal stage. Enzymatic activity analysis of the recombinant enzymes expressed in Escherichia coli showed that FoAK is highly stereo specific for L-arginine versus D-arginine and the apparent Michaelis constant for L-arginine (KmArg) is comparable to that of AKs from a variety of species. This research should enable further investigation of the function as well as in vitro screening for inhibitors of FoAK.

Keywords: Arginine kinase; Frankliniella occidentalis; Heterologous expression; Insecticide target; Kinetic constant.

MeSH terms

  • Animals
  • Arginine Kinase* / biosynthesis
  • Arginine Kinase* / chemistry
  • Arginine Kinase* / genetics
  • Gene Expression*
  • Insect Proteins* / biosynthesis
  • Insect Proteins* / chemistry
  • Insect Proteins* / genetics
  • Introduced Species*
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / genetics
  • Thysanoptera* / enzymology
  • Thysanoptera* / genetics

Substances

  • Insect Proteins
  • Recombinant Proteins
  • Arginine Kinase