Diversity of trypsins in the Mediterranean corn borer Sesamia nonagrioides (Lepidoptera: Noctuidae), revealed by nucleic acid sequences and enzyme purification

Díaz-Mendoza, Mercedes; Ortego, Félix; Lacoba, Mario Garcı́a de; Magaña, C.; Poza, Marta de la; Farinós, Gema P.; Castañera, Pedro; Hernández‐Crespo, Pedro

doi:10.1016/j.ibmb.2005.04.003

Cited by 31 publications

(50 citation statements)

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“…The data on azocasein hydrolysis suggested that adults have trypsin-like enzymes, while PMSF-sensitive enzymes were not found in nymphs. Dıaz-Mendoza et al (2005) reported that E-64 nonspecifically inhibited trypsin-1 enzymes from insects. The more pronounced effect of E-64 on AH activity upon the C-terminal substrate could thus represent the inhibition of cysteine proteinases plus nonspecific inhibition of trypsin-like enzymes.…”

Section: Discussionmentioning

confidence: 99%

Stage-specific gut proteinases of the cotton stainer bug Dysdercus peruvianus: Role in the release of entomotoxic peptides from Canavalia ensiformis urease

Piovesan

Stanisçuaski

Marco-Salvadori

et al. 2008

Insect Biochemistry and Molecular Biology

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Section: Discussionmentioning

confidence: 99%

Stage-specific gut proteinases of the cotton stainer bug Dysdercus peruvianus: Role in the release of entomotoxic peptides from Canavalia ensiformis urease

Piovesan

Stanisçuaski

Marco-Salvadori

et al. 2008

Insect Biochemistry and Molecular Biology

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“…Numerous digestive proteases and amylases have been characterized from many agriculturally important insects (Bown et al, 1998;Gaines et al, 1999;Cristofoletti et al, 2001;Coffeen and Wolpert, 2004;Gruden et al, 2004;Diaz-Mendoza et al, 2005;Zhu et al, 2005). Subsequently, various proteinaceous inhibitors have been identified against these enzymes from different sources (Farmer et al, 1992;Zhao et al, 1996;Koiwa et al, 2000;Moura and Ryan, 2001).…”

Section: Introductionmentioning

confidence: 99%

Cloning and characterization of cDNAS encoding carboxypeptidase-like proteins from the gut of Hessian fly larvae [Mayetiola destructor (Say)]☆

Xiang¹,

Fellers²,

Zhu³

et al. 2006

Insect Biochemistry and Molecular Biology

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“…Other conditions may also cause aggregation of the protein molecules which have amorphous physical appereance. As opposed to gelation, protein aggregation/precipitation is considered to be a phase separation resulting in formation of a solid phase in a solution [2][3]. The aim of this research is to compare the solution, gel and aggregate forms of trypsin obtained in three different environmental conditions using functional and structural means.…”

Section: Introductionmentioning

confidence: 99%

Structural and functional characterization of solution, gel, and aggregated forms of trypsin in organic solvent-assisted and pH-induced phase changes

Ceylan

2014

Turk J Bioch

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Structural and functional characterization of solution, gel, and aggregated forms of trypsin in organic solvent-assisted and pH-induced phase changes [Trpsinin çözelti, jel ve agregat formlarının organik çözgen içeren ve pH-tektiklenmiş faz geçişlerinde yapısal ve fonksiyonel incelenmesi] ABSTRACT Objective: In this study the effect of three different physicochemical parameters on pH-triggered gelation and aggregation of bovine pancreatic trypsin phase changes and phase structural and functional changes in these phase changes in alcohol-water mixtures were studied. Methods: Trypsin gelation times were studied using inverted tube method. Trypsin stability was studied using trypsin enzyme assay. Protein secondary structural changes were monitored using FTIR spectroscopy. Gel and aggregate macrostructures and morphologies were viewed using Scanning Electron Microscopy. Results: The solution phase was observed in the absence of both NaOH and CaCl 2 . The gel phase was observed in the absence of the either. The aggregate phase was observed in the presence of the both agents all depending on trypsin concentrations used. Trypsin stability studies showed that there were a nearly 53 and 32% a specific activity losses after the gelation and aggregation processes. According to FTIR studies β-sheet structure in 1637 cm -1 band disappeared in trypsin gel and trypsin aggregates. Increases in α-helix structure in 1651 cm-1 in trypsin gel and aggregates were observed. Iodoacetamide delayed the gelation and prevented the aggregation indicating the importance of intermolecular disulfides in the both processes. Conclusion: Trypsin gelation was caused by the denaturation of the protein three dimensional structures. The gel and aggregate formation indicates a secondary structural change towards α-helix structure formation at the expense of β-sheet structure and formation of intermolecular disulfide bonds. Key Words: Trypsin, Gelation, Aggregation, FTIR Conflict of Interest: Authors have no conflict of interest. ÖZETAmaç: Bu çalışmada alkol-su karışımlarında pH tarafından tetiklenen sığır pankreatik tripsin jelleşmesi ve agregasyonu faz değişiklikleri üzerine üç değişik fizikokimyasal değişkenin yapısal ve fonksiyonel etkileri çalışılmıştır. Metod: Tripsin jelleşmesi ters tüp metodu ile çalışılmıştır. Tripsin stabilitesi tripsin enzim aktivitesi tayiniyle çalışılmıştır. Protein ikinci yapı değişiklikleri FTIR spektroskopisi metodu ile gözlenmiştir. Jel ve agregat makroyapıları ve morfolojileri taramalı elektron mikroskopisi yöntemiyle incelenmiştir. Bulgular: NaOH ve CaCl 2 eksikliğinde çözelti fazı gözlenmiştir. Jel fazı bu ikisinden birinin eksikliğinde gözlenmiştir. Agregat fazı ise bu iki ajanın birlikte varlığında tripsin konantrasyonuna bağlı olarak gözlenmiştir. Tripsin dayanıklılık çalışmaları jelleşmede yaklaşık olarak 53% ve agregasyonda ise 32% özgül aktivitede dayanıklılık kaybı göstermiştir. FTIR spektroskopisi çalışmalarında tripsin jel ve agregatlarında 1637 cm-1 daki β-yaprağı bantının yok olduğu görülmüştür. 1651 cm-...

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Diversity of trypsins in the Mediterranean corn borer Sesamia nonagrioides (Lepidoptera: Noctuidae), revealed by nucleic acid sequences and enzyme purification

Cited by 31 publications

References 50 publications

Stage-specific gut proteinases of the cotton stainer bug Dysdercus peruvianus: Role in the release of entomotoxic peptides from Canavalia ensiformis urease

Stage-specific gut proteinases of the cotton stainer bug Dysdercus peruvianus: Role in the release of entomotoxic peptides from Canavalia ensiformis urease

Cloning and characterization of cDNAS encoding carboxypeptidase-like proteins from the gut of Hessian fly larvae [Mayetiola destructor (Say)]☆

Structural and functional characterization of solution, gel, and aggregated forms of trypsin in organic solvent-assisted and pH-induced phase changes

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