Preliminary characterisation of digestive proteases of the green mirid, Creontiades dilutus (Hemiptera: Miridae)

Colebatch, Gillian

doi:10.1016/s0965-1748(00)00136-3

Cited by 39 publications

(19 citation statements)

References 31 publications

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“…N-Bnz-R-pna (Eckfeldt et al, 1982), N-Bnz-Y-pna (Colebatch et al, 2001), N-Bnz-FVR-pna (Ashizawa et al, 1998), N-Bnz-PFR-pna (Lai et al, 1991), N-Suc-AAV-pna, and N-Suc-AAPI-pna (Koehl et al, 2003) were all dissolved in DMF, to a 10% final concentration of DMF in the reaction (v/v). Final substrate concentration in the reaction was 1 mM for all except N-Bnz-Y-pna which was 0.5 mM and Z-GGL-pna (Virca et al, 1984) which was 0.2 mM.…”

Section: Protease and Inhibition Assaysmentioning

confidence: 99%

Mating-increases trypsin in female Drosophila hemolymph

Pilpel

Nezer

Applebaum

et al. 2008

Insect Biochemistry and Molecular Biology

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Section: Protease and Inhibition Assaysmentioning

confidence: 99%

Mating-increases trypsin in female Drosophila hemolymph

Pilpel

Nezer

Applebaum

et al. 2008

Insect Biochemistry and Molecular Biology

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“…Esse resultado é um indicativo da presença de cisteíno protease do tipo papaína-like no intestino de A. gemmatalis. Colebatch et al (2001), trabalhando com Figura 4 -Gráfico de Michaelis-Menten da atividade de cisteíno proteases do intestino médio de Anticarsia gemmatalis (Hübner, 1818). Inserção: Gráfico de Lineaweaver-Burk da atividade do extrato enzimático sobre L-BApNA.…”

Section: Methodsunclassified

Caracterização enzimática de isoformas de cisteíno protease de Anticarsia gemmatalis (Hübner, 1818)

et al. 2011

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RESUMOIsoformas de cisteíno protease obtidas do intestino médio de lagartas de 5º instar de Anticarsia gemmatalis (Hübner, 1818) foram caracterizadas. A isoforma solúvel foi chamada de Fração Solúvel enquanto a isoforma ligada à membrana celular, de Fração Insolúvel. As maiores atividades foram observadas em pH 3,6 a 45° C para a Fração Solúvel e pH 4,6 a 50° C para a Fração Insolúvel. Ao analisar o efeito de modificadores químicos, a Fração Solúvel mostrou-se insensível à aprotinina e E-64, porém teve sua atividade aumentada pela adição de EDTA e levemente inibida pela adição de íons Ca 2+ , mostrando se tratar de enzimas independentes de íons metálicos para sua atividade. A Fração Insolúvel também se mostrou insensível à aprotinina, porém teve sua atividade parcialmente inibida por E-64. A adição de EDTA levou a uma redução nos valores de atividade, demonstrando a necessidade de íons metálicos para a atividade dessas enzimas, porém não se trata de enzimas cálcio-dependentes, uma vez que sua atividade foi reduzida com a adição desse íon. Os valores de K M app e V máx app foram, respectivamente, 0,6398 mM e 42,556 nM s -1 para Fração Solúvel e 0,0413 mM e 10,854 nM s -1 para Fração Insolúvel. Esses resultados fornecem evidências da presença de cisteíno protease solúvel e ligada à membrana celular do intestino de lagartas de A. gemmatalis. O conhecimento e a caracterização das principais classes de proteases presentes no trato digestivo da lagarta da soja, bem como a interação dessas enzimas com inibidores de protease têm uma importante consequência aos programas de melhoramento de soja.Termos para indexação: Proteases digestivas, Lepidoptera, controle de pragas, inibição de proteases. ABSTRACTCysteine protease isoforms obtained from the midgut of 5 th instar Anticarsia gemmatalis (Hübner, 1818) larvae were characterized. The soluble enzyme isoform was called Soluble Fraction and cell membrane-bound enzyme isoform, the Insoluble Fraction. The highest activities were observed at pH 3.6 and 45° C for the Soluble Fraction and pH 4.6 and 50° C for the Insoluble Fraction. Analyzing the effect of chemical modifiers, the Soluble Fraction was shown to be insensitive to aprotinin and E-64, although its activity was increased by the addition of EDTA and slightly inhibited by the addition of Ca 2+ , showing to be enzymes independent of metal ions to its activity. The Insoluble Fraction was also insensitive to aprotinin, but its activity was partially inhibited by E-64. The addition of EDTA reduced the activity values, demonstrating the need for metal ions for the activity of these enzymes, but they are not calcium-dependent enzymes, since their activity was reduced with the addition of this ion. The values of K M app and V max app were, respectively, 0.6398 mM and 42.556 nM s -1 for Soluble Fraction and 0.0413 mM and 10.854 nM s -1 for Insoluble Fraction. These results provide evidence for the presence of soluble and cell membrane-bound cysteine proteases in the gut of A. gemmatalis larvae. The knowledge and characterizatio...

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“…Serine proteases similar to those from mammals, specially with respect to the optimum pH, are found as predominant digestive enzymes in a wide variety of insects, as Thysanura (Zinkler & Polzer, 1992), Orthoptera (Lam et al, 1999(Lam et al, , 2000, Hymenoptera (Schumaker et al, 1993), Diptera, (Silva et al, 2006), Lepidoptera (Bernardi et al, 1996;Gatehouse et al, 1999;Novillo et al, 1997) and Hemiptera (Colebatch et al, 2001). Trypsin-like proteases were also found in Coleopteran insects, although be known that these insects have an acid intestinal fluid (Alarcon et al, 2002;Franco et al, 2004;Girard et al, 1998;Purcell et al, 1992;Zhu & Baker, 1999.…”

Section: Serine Proteasesmentioning

confidence: 99%