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Intestinal proteolytic profile changes during larval development of Anticarsia gemmatalis caterpillars.
Arch Insect Biochem Physiol. 2020 Jan; 103(1):e21631.AI

Abstract

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 showed major activities in the late stages of the larval phase. Zymogram analysis and protein identification by liquid chromatography-mass spectrometry indicated serine protease as the main protease class expressed in the fifth instar. These results may shift the focus from the rational development of the protease inhibitor to A. gemmatalis and other Lepidoptera, as the expression of major proteases is not constant.

Authors+Show Affiliations

Department of Biochemistry and Molecular Biology, Laboratory of Enzymology and Biochemistry of Proteins and Peptides, BIOAGRO/INCT-IPP, Universidade Federal de Viçosa, Viçosa, Brazil.Department of Biochemistry and Molecular Biology, Laboratory of Enzymology and Biochemistry of Proteins and Peptides, BIOAGRO/INCT-IPP, Universidade Federal de Viçosa, Viçosa, Brazil. Center of Analysis of Biomolecules, NuBioMol, Universidade Federal de Viçosa, Viçosa, Brazil.Department of Biochemistry and Molecular Biology, Laboratory of Enzymology and Biochemistry of Proteins and Peptides, BIOAGRO/INCT-IPP, Universidade Federal de Viçosa, Viçosa, Brazil.Department of Biochemistry and Molecular Biology, Laboratory of Enzymology and Biochemistry of Proteins and Peptides, BIOAGRO/INCT-IPP, Universidade Federal de Viçosa, Viçosa, Brazil.Department of Biochemistry and Molecular Biology, Laboratory of Enzymology and Biochemistry of Proteins and Peptides, BIOAGRO/INCT-IPP, Universidade Federal de Viçosa, Viçosa, Brazil.Center of Analysis of Biomolecules, NuBioMol, Universidade Federal de Viçosa, Viçosa, Brazil.Department of Entomology, Universidade Federal de Viçosa, Viçosa, Brazil.Department of Biochemistry and Molecular Biology, Laboratory of Enzymology and Biochemistry of Proteins and Peptides, BIOAGRO/INCT-IPP, Universidade Federal de Viçosa, Viçosa, Brazil. Center of Analysis of Biomolecules, NuBioMol, Universidade Federal de Viçosa, Viçosa, Brazil.Department of Biochemistry and Molecular Biology, Laboratory of Enzymology and Biochemistry of Proteins and Peptides, BIOAGRO/INCT-IPP, Universidade Federal de Viçosa, Viçosa, Brazil.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31587381

Citation

da Silva Júnior, Neilier R., et al. "Intestinal Proteolytic Profile Changes During Larval Development of Anticarsia Gemmatalis Caterpillars." Archives of Insect Biochemistry and Physiology, vol. 103, no. 1, 2020, pp. e21631.
da Silva Júnior NR, Vital CE, de Almeida Barros R, et al. Intestinal proteolytic profile changes during larval development of Anticarsia gemmatalis caterpillars. Arch Insect Biochem Physiol. 2020;103(1):e21631.
da Silva Júnior, N. R., Vital, C. E., de Almeida Barros, R., Faustino, V. A., Monteiro, L. P., Barros, E., de Oliveira, E. E., de Oliveira Ramos, H. J., & de Almeida Oliveira, M. G. (2020). Intestinal proteolytic profile changes during larval development of Anticarsia gemmatalis caterpillars. Archives of Insect Biochemistry and Physiology, 103(1), e21631. https://doi.org/10.1002/arch.21631
da Silva Júnior NR, et al. Intestinal Proteolytic Profile Changes During Larval Development of Anticarsia Gemmatalis Caterpillars. Arch Insect Biochem Physiol. 2020;103(1):e21631. PubMed PMID: 31587381.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Intestinal proteolytic profile changes during larval development of Anticarsia gemmatalis caterpillars. AU - da Silva Júnior,Neilier R, AU - Vital,Camilo E, AU - de Almeida Barros,Rafael, AU - Faustino,Verônica A, AU - Monteiro,Luana P, AU - Barros,Edvaldo, AU - de Oliveira,Eugênio E, AU - de Oliveira Ramos,Humberto J, AU - de Almeida Oliveira,Maria G, Y1 - 2019/10/06/ PY - 2019/04/08/received PY - 2019/08/26/revised PY - 2019/09/23/accepted PY - 2019/10/7/pubmed PY - 2020/1/9/medline PY - 2019/10/7/entrez KW - activity KW - intestine KW - plasticity KW - proteases KW - soybean caterpillar SP - e21631 EP - e21631 JF - Archives of insect biochemistry and physiology JO - Arch Insect Biochem Physiol VL - 103 IS - 1 N2 - 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 showed major activities in the late stages of the larval phase. Zymogram analysis and protein identification by liquid chromatography-mass spectrometry indicated serine protease as the main protease class expressed in the fifth instar. These results may shift the focus from the rational development of the protease inhibitor to A. gemmatalis and other Lepidoptera, as the expression of major proteases is not constant. SN - 1520-6327 UR - https://www.unboundmedicine.com/medline/citation/31587381/Intestinal_proteolytic_profile_changes_during_larval_development_of_Anticarsia_gemmatalis_caterpillars_ DB - PRIME DP - Unbound Medicine ER -