Novel Peptidase Kunitz Inhibitor from <i>Platypodium elegans</i> Seeds Is Active against <i>Spodoptera frugiperda</i> Larvae

Ramalho, Suellen Rodrigues; Bezerra, Cézar da Silva; Oliveira, Daniella Gorete Lourenço de; Lima, Letícia Souza; Neto, Simone Maria; Oliveira, Caio Fernando Ramalho de; Verbisck, Newton V.; Macedo, Maria Lı́gia Rodrigues

doi:10.1021/acs.jafc.7b04159

Cited by 21 publications

(6 citation statements)

References 57 publications

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“…Primary metabolism genes such as 4-coumarate-CoA ligaselike 9 and sorbitol dehydrogenase-like were upregulated in H. armigera larvae exposed to SPIs (Table S1). This was consistent with previous work demonstrating that SPI ingestion impacts the regulation of genes involved in carbohydrate and lipid metabolisms in H. armigera, in addition to genes encoding digestive and detoxification enzymes [10,22]. A similar pattern was observed in the generalist herbivore L. migratoria [15].…”

Section: Discussionsupporting

confidence: 92%

Adaptation of Helicoverpa armigera to Soybean Peptidase Inhibitors Is Associated with the Transgenerational Upregulation of Serine Peptidases

Velasquez-Vasconez

Hunt

Dias

et al. 2022

IJMS

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Molecular phenotypes induced by environmental stimuli can be transmitted to offspring through epigenetic inheritance. Using transcriptome profiling, we show that the adaptation of Helicoverpa armigera larvae to soybean peptidase inhibitors (SPIs) is associated with large-scale gene expression changes including the upregulation of genes encoding serine peptidases in the digestive system. Furthermore, approximately 60% of the gene expression changes induced by SPIs persisted in the next generation of larvae fed on SPI-free diets including genes encoding regulatory, oxidoreductase, and protease functions. To investigate the role of epigenetic mechanisms in regulating SPI adaptation, the methylome of the digestive system of first-generation larvae (fed on a diet with and without SPIs) and of the progeny of larvae exposed to SPIs were characterized. A comparative analysis between RNA-seq and Methyl-seq data did not show a direct relationship between differentially methylated and differentially expressed genes, while trypsin and chymotrypsin genes were unmethylated in all treatments. Rather, DNA methylation potential epialleles were associated with transcriptional and translational controls; these may play a regulatory role in the adaptation of H. armigera to SPIs. Altogether, our findings provided insight into the mechanisms of insect adaptation to plant antiherbivore defense proteins and illustrated how large-scale transcriptional reprograming of insect genes can be transmitted across generations.

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Section: Discussionsupporting

confidence: 92%

Adaptation of Helicoverpa armigera to Soybean Peptidase Inhibitors Is Associated with the Transgenerational Upregulation of Serine Peptidases

Velasquez-Vasconez

Hunt

Dias

et al. 2022

IJMS

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“…The noncompetitive mechanism found for the inhibition of ApTI on the trypsin‐like enzymes of A. gemmatalis agrees with other in silico studies carried out with trypsins from other species of the order Lepidoptera (Migliolo et al, 2010; Ramalho et al, 2018), and the reactive (inhibitory) peptide bond was identified in the alpha chain with the participation of the Arg64 residue and amino acid residues around Pro63, Ile65, and Arg66. This bond is at an exactly homologous position to the reactive sites identified in the soybean inhibitor at Arg‐63‐Ile‐64 (McPherson, Daym, & Larson, 2019) in Albizia at Arg‐66‐I1e‐67 (Sharma, Nath, Kumari, & Bhardwaj, 2012) and in Psophocarpus at Arg‐64‐Ser‐65 (Yamamoto, Hara, & Ikenaka, 1983).…”

Section: Discussionsupporting

confidence: 88%

Noncompetitive tight‐binding inhibition ofAnticarsia gemmatalistrypsins byAdenanthera pavoninaprotease inhibitor affects larvae survival

Meriño‐Cabrera

Mendes

Castro

et al. 2020

Arch Insect Biochem Physiol

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The economic loss in soybean crops caused by the Lepidoptera insects has encouraged the search for new strategies to control this pest, which are currently based on synthetic insecticides. This paper evaluated the ability of ApTI (Adenanthera pavonina trypsin inhibitor) to inhibit trypsin-like proteins from Anticarsia gemmatalis by docking, molecular dynamics, and enzymatic and survival assay. The docking and molecular dynamic simulation between trypsin and ApTI were performed using the program CLUSPRO and NAMD, respectively. The inhibitory constant K i and the inhibition type were determined through chromogenic assays. The survival assay of neonatal larvae under treatment with artificial diet supplemented with ApTI was also performed. The ApTI binding site was predicted to block substrate access to trypsin due to four interactions with the enzyme, producing a complex with a surface area of 1,183.7 Å 2 .The kinetic analysis revealed a noncompetitive tightbinding mechanism. The survival curves obtained using Kaplan-Meier estimators indicated that the highest larvae mortality was 60%, using 1.2 mg of ApTI per 100 ml of artificial diet. The in vitro, in vivo, and in silico studies demonstrated that ApTI is a strong noncompetitive inhibitor of trypsin with biotechnological potential for the control of A. gemmatalis insect. K E Y W O R D S binding, insects, Kunitz inhibitor, molecular docking, noncompetitive, trypsin enzyme

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“…The initial high affinity observed between IETI and trypsin was similar to that observed for the trypsin inhibitor from Entada scandens (Lingaraju and Gowda 2008) and followed the pattern observed for PIs with single reactive sites. The experimentally determined dissociation constant (Ki) was 6.2 nM, and the enzyme showed (Macedo et al 2007;Oliveira et al 2012;Ramalho et al 2018). The stability of IETI inhibitory activity was assayed over a range of temperatures, pHs and in the presence of the reducing agent, DTT.…”

Section: Resultsmentioning

confidence: 98%

Biochemical characterization of a Kunitz inhibitor from Inga edulis seeds with antifungal activity against Candida spp.

Dib

Oliveira

et al. 2018

Arch Microbiol

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We describe the characterization of IETI, the first trypsin inhibitor purified from Inga edulis, a tree widely distributed in Brazil. Two-step chromatography was used to purify IETI, a protein composed of a single peptide chain of 19,685.10 Da. Amino-terminal sequencing revealed that IETI shows homology with the Kunitz family, as substantiated by its physicalchemical features, such as its thermal (up to 70 °C) and wide-range pH stability (from 2 to 10), and the value of its dissociation constant (6.2 nM). IETI contains a single reactive site for trypsin, maintained by a disulfide bridge; in the presence of DTT, its inhibitory activity was reduced in a time-and concentration-dependent manner. IETI presented activity against Candida ssp., including C. buinensis and C. tropicalis. IETI inhibitory activity triggered yeast membrane permeability, affecting cell viability, thus providing support for the use of IETI in further studies for the control of fungal infections.

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Novel Peptidase Kunitz Inhibitor from Platypodium elegans Seeds Is Active against Spodoptera frugiperda Larvae

Cited by 21 publications

References 57 publications

Adaptation of Helicoverpa armigera to Soybean Peptidase Inhibitors Is Associated with the Transgenerational Upregulation of Serine Peptidases

Adaptation of Helicoverpa armigera to Soybean Peptidase Inhibitors Is Associated with the Transgenerational Upregulation of Serine Peptidases

Noncompetitive tight‐binding inhibition ofAnticarsia gemmatalistrypsins byAdenanthera pavoninaprotease inhibitor affects larvae survival

Biochemical characterization of a Kunitz inhibitor from Inga edulis seeds with antifungal activity against Candida spp.

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