Screening of Vip3Aa60 and Vip3Ad5 and characterization of their binding to Spodoptera exigua midguts

Pan, Zhiyong; Xu, Liang; Zhang, Jing; Liu, Bo; Chen, Qing‐Xi; Gao, Huan-Juan; Chen, Zheng; Zhu, Yujing; Guan, Xiong

doi:10.1016/j.procbio.2017.06.023

“…Previous study indicated that both Vip3Aa and Vip3Ad could bind to S. exigua BBMV, while the binding amount of Vip3Aa was more than Vip3Ad, which might be one reason for the difference in insecticidal activities between Vip3Aa and Vip3Ad (Pan et al, 2017). However, considering that Vip3Ad was nontoxic at all to S. exigua, we believed there were other factors resulting in the total inactivity of Vip3Ad.…”

Section: Discussionmentioning

confidence: 80%

“…Protein concentration was determined using a BCA Protein Assay Kit (Beyotime, China). Trypsin or S. exigua midgut juice was used to produce Vip3Aa and Vip3Ad activatedtoxins as previous described (Pan et al, 2017).…”

Section: Insectsmentioning

confidence: 99%

“…The insecticidal activities of Vip3Aa and Vip3Ad against rst-instar larvae of S. exigua were estimated according to Pan et al (2017). Six concentrations of toxins were set up, and sodium carbonate buffer (50 mM, pH 9.5) was served as a negative control.…”

Section: Bioassaysmentioning

confidence: 99%

“…Previous study demonstrated that Vip3Aa and Vip3Ad showed signi cant difference in insecticidal activities against lepidopteran pest Spodoptera exigua, while the molecular mechanism was still unclear (Pan et al, 2017). Here we found that both of Vip3Aa and Vip3Ad could be cleaved into activated-toxins by Spodoptera exigua midgut protease and their cleavage sites were the same.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Stability is essential for insecticidal activity of Vip3Aa toxin against Spodoptera exigua

Fu

¹

,

Liu

²

,

Zheng

³

et al. 2022

Preprint

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Vegetative insecticidal proteins 3A (Vip3A) were important insecticidal proteins for control of lepidopteran pests. Previous study demonstrated that Vip3Aa and Vip3Ad showed significant difference in insecticidal activities against Spodoptera exigua, while the molecular mechanism remained ambiguous. Here we demonstrated that the difference in insecticidal activities between Vip3Aa and Vip3Ad might be caused by the difference in stability of Vip3Aa and Vip3Ad in S. exigua midgut protease. Vip3Aa was quite stable while Vip3Ad could be further degraded. Molecular dynamics simulation revealed that Vip3Aa was more stable than Vip3Ad, with smaller RMSD and RMSF value. Amino acid sequence alignment indicated that three were three extra prolines (P591, P605 and P779) located on Vip3Aa. We further identified that residue P591 played a crucial role on stability and insecticidal activity of Vip3Aa. Taken together, our study demonstrated that the stability was essential for the insecticidal activity of Vip3A toxins, which might provide new insight into the action mode of Vip3A toxins and contribute to the design Vip3A variants with improved stability and insecticidal activity.

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“…Pull-down experiments were also performed using Cathepsin Bcoupled Sepharose 4B beads with the purified recombinant Cry41related toxin or bovine serum albumin (BSA) as a control. Cry41related toxin or the BSA solution was subjected to the Cathepsin Bcoupled Sepharose 4B column, the flow-through from the column and the proteins bound to the Cathepsin B column were analyzed by Western blot according to the method described by Pan et al 26 Effect of Temperature and pH on Cathepsin B Activity and Stability. The assay for Cathepsin B activity was performed according to the method described by Abood et al 27 Briefly, the optimum temperature of Cathepsin B activity was determined at various temperatures (20−50 °C) for 1 h using BSA as the substrate under standard assay conditions.…”

Section: T H I S C O N T E N T I S O N L Y L I C E N S E D F O R C O ...mentioning

confidence: 99%

Possible Insecticidal Mechanism of Cry41-Related Toxin against Myzus persicae by Enhancing Cathepsin B Activity

Zhao

¹

,

Zhang

²

,

Fu

³

et al. 2020

J. Agric. Food Chem.

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Cry toxins produced by Bacillus thuringiensis are well known for their high insecticidal activities against Lepidoptera, Diptera, and Coleoptera; however, their activities against Aphididae are very low. Recently, it has been reported that a Cry41-related toxin exhibited moderate activity against the aphid Myzus persicae, and thus, it is highly desirable to uncover its unique mechanism. In this paper, we report that Cathepsin B, calcium-transporting ATPase, and symbiotic bacterial-associated protein ATP-dependent-6-phosphofructokinase were pulled down from the homogenate of M. persicae as unique proteins that possibly bound to Cry41related toxin. Cathepsin B has been reported to cleave and inactivate antiapoptotic proteins and plays a role in caspase-initiated apoptotic cascades. In this study, Cathepsin B was expressed in Escherichia coli and purified, and in vitro interaction between recombinant Cathepsin B and Cry41-related toxin was demonstrated. Interestingly, we found that addition of Cry41-related toxin obviously enhanced Cathepsin B activity. We propose a model for the mechanism of Cry41-related toxin as follows: Cry41-related toxin enters the aphid cells and enhances Cathepsin B activity, resulting in acceleration of apoptosis of aphid cells.

show abstract

Stability is essential for insecticidal activity of Vip3Aa toxin against Spodoptera exigua

Fu

¹

,

Xu

²

,

Zheng

³

et al. 2022

View full text Add to dashboard Cite

Vegetative insecticidal proteins 3A (Vip3A) were important insecticidal proteins for control of lepidopteran pests. Previous study demonstrated that Vip3Aa and Vip3Ad showed significant difference in insecticidal activities against Spodoptera exigua, while the molecular mechanism remained ambiguous. Here we demonstrated that the difference in insecticidal activities between Vip3Aa and Vip3Ad might be caused by the difference in stability of Vip3Aa and Vip3Ad in S. exigua midgut protease. Vip3Aa was quite stable while Vip3Ad could be further degraded. Molecular dynamics simulation revealed that Vip3Aa was more stable than Vip3Ad, with smaller RMSD and RMSF value. Amino acid sequence alignment indicated that three were three extra prolines (P591, P605 and P779) located on Vip3Aa. We further identified that residue P591 played a crucial role on stability and insecticidal activity of Vip3Aa. Taken together, our study demonstrated that the stability was essential for the insecticidal activity of Vip3A toxins, which might provide new insight into the action mode of Vip3A toxins and contribute to the design Vip3A variants with improved stability and insecticidal activity.

show abstract

Screening of Vip3Aa60 and Vip3Ad5 and characterization of their binding to Spodoptera exigua midguts

Cited by 3 publications

References 38 publications

Stability is essential for insecticidal activity of Vip3Aa toxin against Spodoptera exigua

Stability is essential for insecticidal activity of Vip3Aa toxin against Spodoptera exigua

Possible Insecticidal Mechanism of Cry41-Related Toxin against Myzus persicae by Enhancing Cathepsin B Activity

Stability is essential for insecticidal activity of Vip3Aa toxin against Spodoptera exigua

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