Characterization of Trypsin Like Protease from Helicoverpa armigera (Hubner) and Its Potential Inhibitors

Grover, Sheetanshu; Kaur, Sarvjeet; Gupta, Anil Kumar; Taggar, Gaurav Kumar; Kaur, Jagmeet

doi:10.1007/s40011-016-0732-0

Cited by 11 publications

(7 citation statements)

References 26 publications

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“…The fact that trypsin‐ like and chymotrypsin‐ like increased significantly from third to fourth and from fourth to fifth instar corroborates and confirms the results obtained by specific inhibitors. The results also show that trypsin‐ like enzymes are more efficient than chymotrypsin‐ like enzymes in the fifth instar, corroborating with previous studies that demonstrated the prevalence of this enzyme group in the fifth and sixth instar of Lepidoptera larvae (Sharifloo, Zibaee, Sendi, & Talebi, ; Sharath Chandra et al, ; Grover, Kaur, Gupta, Taggar, & Kaur, ). The activity of cysteine proteases was determined by means of the substrate PFLNA, and the results showed that the specific activity of this sub‐subclass decreased with time.…”

Section: Resultssupporting

confidence: 90%

Intestinal proteolytic profile changes during larval development of Anticarsia gemmatalis caterpillars

Junior

Vital

Barros

et al. 2019

Arch Insect Biochem Physiol

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Soybean is one of most consumed and produced grains in the world, and Anticarsia gemmatalis is a pest that causes great damage to this crop due to severe defoliation during its larval phase. Plants have mechanisms that lead to the inhibition of proteases in the intestine of these herbivores, hampering their development. Understanding this complex protease inhibitor is important for pest control. The objective of this study was to evaluate the enzymatic profiles of the intestinal proteases of the soybean caterpillar at different instars. For this, the proteolytic profile of the gut in the third, fourth, and fifth instars were analyzed.Irreversible inhibitors of proteases were separately incubated with A. gemmatalis enzyme extracts at the third, fourth, and fifth instar to assess the contribution of these proteases to total proteolytic activity. The enzymatic extracts were also evaluated with specific substrates to confirm changes in the specific activities of trypsin-like, chymotrypsin-like, and cysteine proteases at different instars. The results showed that the protease profile of A.gemmatalis gut changes throughout its larval development.The activity of cysteine proteases was more intense in the first instar. On the contrary, the serine proteases The protease profile of the gut of A. gemmatalis changes throughout larval development. In the third instar, cysteine protease activity is predominant over others. In the fourth and fifth instars, proteolytic activity is mainly due to serine proteases. This difference in activity in the different instars of A. gemmatalis suggests that the expression of the genes coding for proteases also changes throughout the development of the insect. Using serine PIs is a good strategy for controlling insect pests, as these are the main proteolytic enzymes of Lepidoptera. However, the presence of other important protease subclasses in insect digestibility suggests that a combination of serine and cysteine PIs may be promising, aiming to compromise larval development at different instars.

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Section: Resultssupporting

confidence: 90%

Intestinal proteolytic profile changes during larval development of Anticarsia gemmatalis caterpillars

Junior

Vital

Barros

et al. 2019

Arch Insect Biochem Physiol

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“…Trypsin-like protease belongs to the serine protease family. It is responsible for various physiological functions, including protein digestion, protein absorption, and immune response in insects [ 36 ]. Its expression was up-regulated by a factor of 2.18.…”

Section: Discussionmentioning

confidence: 99%

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

Feng

Wei

et al. 2017

Toxins

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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.

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“…Our results showed an enzyme by molecular mass of 25 kDa, optimal pH of 8 and optimal temperature of 40°C. In lepidopterans, the purified trypsins have shown molecular mass of 24.6 kDa from Ostrinia nubilalis Hubner (Lepidoptera: Crambidae), to 24 kDa in Spodoptera littoralis Boisduval (Lepidoptera: Noctuidae), then 28.7 in Diatraea saccharalis Fabricius (Lepidoptera: Crambidae), then 24–27 kDa in Sesamia nonagrioides Lefebvre (Lepidoptera: Noctuidae), Helicoverpa armigera Hubner (Lepidoptera: Noctuidae), further 35.8 kDa in Ectomyelois ceratoniae Walker (Lepidoptera: Pyralidae) and finally 18.8 kDa in H. armigera (Bernardi et al , 1996; Marchetti et al , 1998; Novillo et al , 1999; Telang et al , 2005; Lopes et al , 2006; Ranjbar et al , 2014; Grover et al , 2016). Although two exceptions have been reported in the molecular mass of two trypsins from Anticarsia gemmatalis Hubner (Lepidoptera: Noctuidae) and Naranga aenescens Moore (Lepidoptera: Noctuidae) as 66–91 and 88.5 kDa (Oliveira et al , 2005; Zibaee et al , 2011).…”

Section: Discussionmentioning

confidence: 99%

Biochemical characterization a digestive trypsin in the midgut of large cabbage white butterfly,Pieris brassicaeL. (Lepidoptera: Pieridae)

Sharifloo

Zibaee

Sendi

et al. 2017

Bull. Entomol. Res.

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A comprehensive study on digestive trypsin was undertaken in the larval midgut of Pieris brassicae L. Results of enzymatic compartmentalization showed a significantly higher activity of crude trypsin in the anterior larval midgut rather than posterior-midgut. Using Diethylaminoethyl cellulose fast flow column chromatography a purified trypsin was obtained by specific activity of 21 U mg-1 protein, recovery of 22%, purification fold of 28-fold and molecular weight of 25 kDa. This purified enzyme showed the highest activity at pH 8 and the corresponding temperature of 40°C. However, the specific inhibitors used including 4-(2-Aminoethyl) benzenesulfonyl fluroride hydrochloride, N-p-Tosyl-L-lysine methyl ester hydrochloride and Soybean Trypsin Inhibitor significantly lowered the activity of the purified enzyme in vitro. Moreover, the activity of trypsin and likewise the nutritional indices were significantly altered in the larval midgut feeding upon the leaves treated by 1 mM concentration of each inhibitor in comparison with control. Determination of enzymatic characteristics of insect trypsins is crucial in paving the path for controlling pests by potential natural compounds via transgenic plants.

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Characterization of Trypsin Like Protease from Helicoverpa armigera (Hubner) and Its Potential Inhibitors

Cited by 11 publications

References 26 publications

Intestinal proteolytic profile changes during larval development of Anticarsia gemmatalis caterpillars

Intestinal proteolytic profile changes during larval development of Anticarsia gemmatalis caterpillars

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

Biochemical characterization a digestive trypsin in the midgut of large cabbage white butterfly,Pieris brassicaeL. (Lepidoptera: Pieridae)

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