Structural and mechanistic insights into a novel non-competitive Kunitz trypsin inhibitor from Adenanthera pavonina L. seeds with double activity toward serine- and cysteine-proteinases

Migliolo, Ludovico; Oliveira, Adeliana S.; Santos, Elizeu A.; Franco, Octávio L.; Sales, Maurı́cio P.

doi:10.1016/j.jmgm.2010.05.006

Cited by 50 publications

(38 citation statements)

References 60 publications

(102 reference statements)

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“…In general, Fabaceae seeds have high levels of protease inhibitors and these molecules have been purifi ed from several legume species (Gomes et al 2005, Migliolo et al 2010, Paula et al 2012, Chevreuil et al 2014. For example, in Pithecelobium dumosum, two trypsin inhibitors were purifi ed (Oliveira et al 2007) and in Poincianella pyramidalis seeds, a Kunitz inhibitor with insecticide effect was isolated (Guimarães et al 2015).…”

Section: Discussionmentioning

confidence: 99%

“…Because of the versatility in inhibiting different enzymes, bifunctional inhibitors have been investigated to understand their mechanistic inhibitory strategies, in order to elucidate their interaction with their target enzymes (Migliolo et al 2010) and their potential applications as therapeutic drugs to treat cancer and to control pests and crop diseases in transgenic plants (Pandey et al 2007, Mosolov & Valueva 2008, Paula et al 2012, Cruz-Silva et al 2013.…”

Section: Discussionmentioning

confidence: 99%

“…Of these, the serine protease inhibitor (serpins) and cysteine protease inhibitors (cystatins) are the most widely studied (Oliveira et al 2003, Oliveira et al 2007, Volpicella et al 2011, Fluhr et al 2012, Chevreuil et al 2014. Some protease inhibitors are called bifunctional because they inhibit more than one mechanistic class of proteases (Migliolo et al 2010).…”

Section: Introductionmentioning

confidence: 99%

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Partial purification of trypsin/papain inhibitors from Hymenaea courbaril L. seeds and antibacterial effect of protein fractions

Brito

Melo

Alves

et al. 2016

Hoehnea

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-(Partial purification of trypsin/papain inhibitors from Hymenaea courbaril L. seeds and antibacterial effect of protein fractions). The crude extract and protein fractions of Hymenaea courbaril L. seeds were investigated for the presence of trypsin and papain inhibitors and antimicrobial activity against Vibrio parahaemolyticus, Staphylococcus aureus, and Escherichia coli. Protein fractions were obtained from the crude extract after precipitation with ammonium sulfate into three saturation ranges (0-30%, 30-60%, and 60-90%), called Hc030, Hc3060, and Hc6090, respectively. The crude extract and protein fractions inhibited trypsin and papain activity, but to different degrees. Antimicrobial activity was observed in Hc030 and Hc3060 fractions, but only against V. parahaemolyticus.The inhibitor isolated from the Hc3060 fraction was more effective in inhibiting trypsin (100% inhibition) than papain (54% inhibition), and showed an apparent molecular mass of 20 kDa. This study shows that H. courbaril seeds contain proteins with protease-inhibiting and antibacterial activity, indicating that this species is a source of bioactive compounds. Keywords: antimicrobial, cystatin, Fabaceae, protease RESUMO -(Purificação parcial de um inibidor de tripsina/papaína de sementes de Hymenaea courbaril L. e atividade antibacteriana de suas frações proteicas). O extrato bruto e frações proteicas de sementes de Hymenaea courbaril foram investigados para a presença de inibidores de tripsina e papaína e atividade antimicrobiana contra Vibrio parahaemolyticus, Staphylococcus aureus e Escherichia coli. As frações foram obtidas após precipitação do extrato bruto com sulfato de amônio em três faixas de saturação (0-30%, 30-60% e 60-90%) e denominadas Hc030, Hc3060 e Hc6090, respectivamente. O extrato bruto e as frações apresentaram diferente especificidade em inibir a atividade da tripsina e papaína. A atividade antimicrobiana foi observada nas frações Hc030 e Hc3060 e somente contra V. parahaemolyticus. O inibidor isolado a partir de Hc3060 foi mais eficiente na inibição da tripsina (100% de inibição) do que da papaína (54% de inibição), e mostrou uma massa molecular aparente de 20 kDa. Este estudo mostra que as sementes de H. courbaril possuem inibidores de proteases e proteínas com atividade antibacteriana, indicando que a espécie é uma fonte de compostos bioativos.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Partial purification of trypsin/papain inhibitors from Hymenaea courbaril L. seeds and antibacterial effect of protein fractions

Brito

Melo

Alves

et al. 2016

Hoehnea

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“…Because of this structural feature, they are known as one head inhibitors ("single headed") (Ryan, 1990). On the other hand, few representants of this family were characterized as inhibitors that have two sites for two different enzymes (Migliolo et al, 2010;Valueva et al, 1999;Bösterling & Quast, 1981). Arginine and Lysine residues are part of reactive sites of the inhibitors (Iwanaga et al, 2005).…”

Section: Serine Protease Inhibitorsmentioning

confidence: 99%

“…ApTKI was a strong non-competitive inhibitor of trypsin and moderate noncompetitive inhibitor to papain. Different from PjTKI, that was incapable of simultaneous inhibition of trypsin and papain, the interaction sites of the ApTKI did not overlap, and it formed a ternary complex that was observed through in vitro and in silico methods (Macedo et al, 2004;Prabhu & Pattabiraman, 1980;Migliolo et al, 2010).…”

mentioning

confidence: 99%

Affinity Chromatography as a Key Tool to Purify Protein Protease Inhibitors from Plants

Santos¹,

Oliveira²,

Rabêlo³

et al. 2012

Affinity Chromatography

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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

show abstract

Structural and mechanistic insights into a novel non-competitive Kunitz trypsin inhibitor from Adenanthera pavonina L. seeds with double activity toward serine- and cysteine-proteinases

Cited by 50 publications

References 60 publications

Partial purification of trypsin/papain inhibitors from Hymenaea courbaril L. seeds and antibacterial effect of protein fractions

Partial purification of trypsin/papain inhibitors from Hymenaea courbaril L. seeds and antibacterial effect of protein fractions

Affinity Chromatography as a Key Tool to Purify Protein Protease Inhibitors from Plants

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

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