Comparative Proteomic Analysis of the Effect of Periplocoside P from Periploca sepium on Brush Border Membrane Vesicles in Midgut Epithelium of Mythimna separata Larvae

Abstract

Periplocoside P (PSP), a novel compound isolated from Periploca sepium Bunge, possesses insecticidal activity against some lepidopterans, such as Mythimna separata. In M. separata, the brush border membrane vesicles of the midgut epithelium are the initial site of action of periplocosides. We conducted two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time of flight/time of flight mass spectrometry analysis to analyze differentially expressed proteins (DEPs) from periplocoside P (PSP)-treated M. separata. We successfully isolated seven up-regulated and three down-regulated DEPs that have been previously identified, as well as a novel DEP. The DEPs are implicated in protein degradation, transporter, folding, and synthesis, and in juvenile hormone biosynthesis. DEPs involved in the oxidative phosphorylation energy metabolism pathway are enriched. Through real-time polymerase chain reaction assay, we confirmed that vma1 expression is significantly up-regulated expression levels in PSP-treated M. separata larvae. Enzymology validation further indicated that PSP can significantly inhibit V-type ATPase activity in a concentration-dependent manner. Given these results, we speculate that in M. separata, the V-type ATPase A subunit in the midgut epithelium is the putative target binding site of periplocosides. This finding provides preliminary evidence for the mode of action of periplocosides.

Periplocoside P affects synaptic transmission at the neuromuscular junction and reduces synaptic excitability in Drosophila melanogaster by inhibiting V‐ATPase

“…8 Quantitative differential proteomics studies have demonstrated that PSP treatment markedly upregulates the expression of V-type adenosine triphosphatase (V-ATPase) in M. separata larvae. 9,10 Enzymatic validation further confirmed the concentrationdependent inhibitory effect of PSP on V-ATPase activity. 9,10 Based on the findings mentioned above, we hypothesized that V-ATPase serves as a target for the insecticidal activity of PSP.…”

“…9,10 Enzymatic validation further confirmed the concentrationdependent inhibitory effect of PSP on V-ATPase activity. 9,10 Based on the findings mentioned above, we hypothesized that V-ATPase serves as a target for the insecticidal activity of PSP.…”

Genus Periploca (Apocynaceae): A Review of Its Classification, Phytochemistry, Biological Activities and Toxicology

“…They speculated that periplocoside P adjusted the transport mechanisms at the apical membrane of the midgut epithelial cells by inhibiting the V-type H + ATPase [194]. In addition, the V-type ATPase A subunit in the midgut epithelium might be regarded as the target binding site of periplocosides, which provided preliminary evidence for the mode of action of periplocosides [195]. Furthermore, aminopeptidases N and N3 of the midgut epithelium cells of M. separata larvae were extrapolated as potential targets of periplocoside E ( 47 ) by affinity chromatography [196].…”

Identification and mechanism of insecticidal periplocosides from the root bark of Periploca sepium Bunge

“…23 Additionally, further elaboration of these periplocosides' mode action has been conducted, with results showing that periplocosides could activate protease from the midgut of M. separata after being fed to pests. [24][25][26][27] Notably, it has also been reported that the crude fraction F3-28 made of various periplocosides form PSRB could not only activate protease but it also activated tryptase in vivo. 28 These studies indicated periplocosides could affect mainly digestive enzymes in vivo.…”

“…Periplocoside X (PSX) was used as a botanical pesticide against workers of the red imported fire ants (LD 50 = 0.116 mg/mL) 23 . Additionally, further elaboration of these periplocosides' mode action has been conducted, with results showing that periplocosides could activate protease from the midgut of M. separata after being fed to pests 24–27 . Notably, it has also been reported that the crude fraction F3‐28 made of various periplocosides form PSRB could not only activate protease but it also activated tryptase in vivo 28 .…”