Isolation and characterization of two digestive trypsin-like proteinases from larvae of the stalk corn borer, Sesamia nonagrioides

Novillo, Concepción; Castañera, Pedro; Ortego, Félix

doi:10.1016/s0965-1748(98)00121-0

Cited by 36 publications

(66 citation statements)

References 23 publications

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“…In contrast to many other insects (Milne and Kaplan, 1993;Valaitis, 1995;Ortego et al, 1996;Novillo et al, 1999;Wagner et al, 2002) we observed low specific activities for trypsin-like and chymotrypsin-like enzymes, we have found no activity for elastolytic enzymes. The rate of breakdown of natural substrate hemoglobin by non specific proteases, probably cathepsin D-like enzymes (Lemos and Terra, 1991), was also low.…”

Section: Discussioncontrasting

confidence: 99%

“…Influence of pH on peptidolytic activity of housefly larvae enzymes is similar to many other insect proteinases. Most serine and metallo enzymes of larval origin have pH optimum in slightly alkalic region, pH 8-9 (Lemos and Terra, 1992;Johnson et al, 1995;Lam et al, 1999;Novillo et al, 1999;Bozic et al, 2003). This pH was observed also in our case for hydrolysis of both chromogenic substrates and hemoglobin, too.…”

Section: Discussionmentioning

confidence: 99%

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Peptidolytic enzymes in different larval stadium of housefly Musca domestica

Blahovec¹,

Kostecká²,

Kočišová³

2006

Vet. Med.

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Four classes of peptidolytic enzymes were described in insects. Many authors have found predominant activity belonging to trypsin-like and chymotrypsin-like activity. By the use specific chromogenic substrates and hemoglobin we have determined enzyme activity in three stages of larval development of housefly. In contrast to above mentioned data we have found, that major part of peptidolytic activity in this insect is of aminopeptidase nature. Other observed peptidolytic activity formed only minority part. Apparently the highest activities to all examined substrates were found in first larval stadium of housefly. Inhibitory studies by class specific inhibitors and influence of metal ions and chelating agent on enzyme activity have shown, that aminopeptidase-like enzymes belong to metalloproteinase group.

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Section: Discussioncontrasting

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Peptidolytic enzymes in different larval stadium of housefly Musca domestica

Blahovec¹,

Kostecká²,

Kočišová³

2006

Vet. Med.

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“…TLCK, an irreversible inhibitor of trypsin-like serine proteases, alkylates an histidine residue close to the reactive serine of trypsins . The membrane-bound proteases of A. gemmatalis were inactivated not only by PMSF, but also by TLCK ( Table 1) that inhibits trypsins (Bernardi et al, 1996;Gatehouse et al, 1999;Novillo et al, 1999;Zhu & Baker, 1999;Lam et al, 2000).…”

Section: Inhibition Studiesmentioning

confidence: 99%

“…The digestive activity of trypsin-like enzymes from insect midgut were studied and characterized in Auchenorrhyncha (Foissac et al, 2002), Blattodea (Elpidina et al, 2001;Marana et al, 2002;Lopes & Terra, 2003), Coleoptera (Zhu & Baker, 1999;Wagner et al, 2002), Diptera (Gorman & Paskewitz, 2001;Muharsini et al, 2001) and Lepidoptera (Lee & Anstee, 1995;Valaitis, 1995;Novillo et al, 1999).…”

Section: Introductionmentioning

confidence: 99%

Trypsin-like activity of membrane-bound midgut proteases from Anticarsia gemmatalis (Lepidoptera: Noctuidae)

Xavier¹,

Oliveira²,

Guedes³

et al. 2005

Eur. J. Entomol.

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Abstract. Membrane-bound proteases from preparations of the midgut of 5 th instar velvetbean caterpillars, Anticarsia gemmatalis (Hübner) were obtained by resuspension of the pellet obtained after 100,000 g centrifugation. As expected of trypsin-like proteases, they hydrolyzed casein and the synthetic substrates N--benzoyl-L-Arg-p-nitroanilidine (L-BApNA) and N--p-tosyl-L-Arg methyl ester (L-TAME). Higher activities were observed at 50°C, and at pH 8.5 and 8.0 for both synthetic substrates L-BApNA and L-TAME. The membrane-bound proteases were inhibited by EDTA, phenylmethan sulphonyl fluoride (PMSF), tosyl-L-lysine chloromethyl ketone (TLCK), benzamidine and aprotinin. TLCK and benzamidine were particularly active inhibitors. The KM-values obtained were 0.23 mM for L-BApNA and 92.5 µM for L-TAME. These results provide evidence for the presence of membranebound trypsin-like proteases in the midgut of the velvetbean caterpillar, a key soybean pest in warm climates. The interaction between A. gemmatalis digestive proteases and soybean protease inhibitors has potentially important consequences for soybean breeding programs.

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“…As a consequence, a wide range of investigations have been focused on the characterisation and identification of different target proteinases in a variety of insect orders: Lepidoptera, such as Agrotis (Purcell et al, 1992), Cydia (Christeller et al, 1992), Spodoptera (Lee and Anstee, 1995), Lymantria (Valaitis, 1995), Heliothis (Johnston et al, 1995), Ostrinia (Bernardi et al, 1996), Bombyx (Nobuyasu and Yamashita, 1997), Sesamia (Novillo et al, 1999), Pieris (Broadway, 1995); in Coleoptera, such as Adalia (Murdock et al, 1987;Walker et al, 1998), Tenebrio (Dadd, 1956;Jang et al, 1998) and Tribolium (BlancoLabra et al, 1996); in Diptera, such as Aedes (Fisk, 1950) and Aenopheles (Berner et al, 1983;Vizioli et al, 2001); in Hemiptera, such as Dysdercus (Khan and Ford, 1962); and in Orthoptera, such as Locusta (Khan, 1963). The potential damage of protein-inhibitors on the digestive proteinases of beneficial insects has received less attention.…”

Section: Introductionmentioning

confidence: 99%

Expression profile of water-soluble proteinases during ontogenesis of Megachile rotundata: an electrophoretic investigation

et al. 2004

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-Variations in proteinase activity pattern in larva, pupa and imago of the solitary bee Megachile rotundata are described. Extraction of insect homogenates under mild conditions was followed by the electrophoretic separation of the protein extract in polyacrylamide gels, precast with either gelatine or pollen protein extracts. In these conditions, twelve distinct proteinases were detectable in the pooled I-IV instar larvae, six in the pollen-eating V instar, two in the mature V instar, none in the diapausing V instar, none in the pupa, and two in the imago. Some of the detected proteinases were able to digest the protein mixture extracted from the pollen provisions. Some proteinases were insect specific since they were not detectable in pollen provisions extract. The enzymologic properties of the major proteolytic band suggest its serine-proteinase nature.solitary bee / Megachile rotundata / development / proteinase / zymography

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Isolation and characterization of two digestive trypsin-like proteinases from larvae of the stalk corn borer, Sesamia nonagrioides

Cited by 36 publications

References 23 publications

Peptidolytic enzymes in different larval stadium of housefly Musca domestica

Peptidolytic enzymes in different larval stadium of housefly Musca domestica

Trypsin-like activity of membrane-bound midgut proteases from Anticarsia gemmatalis (Lepidoptera: Noctuidae)

Expression profile of water-soluble proteinases during ontogenesis of Megachile rotundata: an electrophoretic investigation

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