Molecular characterization of a voltage-gated calcium channel and its potential role in the acaricidal action of scopoletin against Tetranychus cinnabarinus.

The carmine spider mite, Tetranychus cinnabarinus (Boisduval), is a polyphagous agricultural pest with an extensive host plant range. Scopoletin is a promising acaricidal compound whose acaricidal mechanism may occur by disrupting intracellular Ca2+ homeostasis and calcium signaling pathways. However, the underlying mechanism of scopoletin for specific target locations of T. cinnabarinus remains unclear. In this study, a full-length cDNA of the L-type voltage-gated calcium channel (TcLTCC) subunit gene from T. cinnabarinus was cloned and characterized. The expression pattern of the TcLTCC gene in all developmental stages of T. cinnabarinus was analyzed. The gene was highly expressed in larval and nymphal stages and was significantly upregulated after treatment with scopoletin. Knocking down the TcLTCC transcript reduced the sensitivity of T. cinnabarinus to scopoletin. Homology modeling and molecular docking were also conducted. The interaction between scopoletin and TcLTCC showed that scopoletin inserted into the cavity bound to the site of the TcLTCC protein by the driving force of hydrogen bonding. This study provides insights into the mechanism by which scopoletin interacts with TcLTCC. Results can improve the understanding of the toxicity of scopoletin to T. cinnabarinus and provide valuable information for the design of new LTCC inhibitors.

3D-QSAR and Molecular Docking Studies on the TcPMCA1-Mediated Detoxification of Scopoletin and Coumarin Derivatives.

The carmine spider mite, Tetranychus cinnabarinus (Boisduval), is an economically important agricultural pest that is difficult to prevent and control. Scopoletin is a botanical coumarin derivative that targets Ca2+-ATPase to exert a strong acaricidal effect on carmine spider mites. In this study, the full-length cDNA sequence of a plasma membrane Ca2+-ATPase 1 gene (TcPMCA1) was cloned. The sequence contains an open reading frame of 3750 bp and encodes a putative protein of 1249 amino acids. The effects of scopoletin on TcPMCA1 expression were investigated. TcPMCA1 was significantly upregulated after it was exposed to 10%, 30%, and 50% of the lethal concentration of scopoletin. Homology modeling, molecular docking, and three-dimensional quantitative structure-activity relationships were then studied to explore the relationship between scopoletin structure and TcPMCA1-inhibiting activity of scopoletin and other 30 coumarin derivatives. Results showed that scopoletin inserts into the binding cavity and interacts with amino acid residues at the binding site of the TcPMCA1 protein through the driving forces of hydrogen bonds. Furthermore, CoMFA (comparative molecular field analysis)- and CoMSIA (comparative molecular similarity index analysis)-derived models showed that the steric and H-bond fields of these compounds exert important influences on the activities of the coumarin compounds.Notably, the C3, C6, and C7 positions in the skeletal structure of the coumarins are the most suitable active sites. This work provides insights into the mechanism underlying the interaction of scopoletin with TcPMCA1. The present results can improve the understanding on plasma membrane Ca2+-ATPase-mediated (PMCA-mediated) detoxification of scopoletin and coumarin derivatives in T. cinnabarinus, as well as provide valuable information for the design of novel PMCA-inhibiting acaricides.