Expression profile of water-soluble proteinases during ontogenesis of <i>Megachile rotundata</i>: an electrophoretic investigation

Felicioli, Antonio; Donadio, Elena; Balestreri, Ettore; Montagnoli, Giorgio; Felicioli, Romano; Podestà, Adriano

doi:10.1051/apido:2004064

Cited by 9 publications

(9 citation statements)

References 44 publications

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“…In the late phase of diapause, the observed increase in proteolytic activity levels in both sexes, in particular in hatching individuals, could be related to the insects' preparations for active life and food intake. A similar increase in proteolytic activity levels was observed after hatching in other insects, including M. rotundata and Bombyx mori (Indrasith et al, 1988;Ikeda et al, 1990;Felicioli et al, 2004). The substrate preferences of the analysed enzymes suggest there are sex-related differences.…”

Section: Resultssupporting

confidence: 65%

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The Proteolytic Activity of Diapausing and Newly Hatched Red Mason Bees (Osmia Rufa: Megachilidae)

Zaobidna

Dmochowska

Frączek

et al. 2014

Journal of Apicultural Science

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a b s t r a c t osmia rufa is a solitary bee that is used commercially for pollinating crops. The bee enters obligatory diapause as an imago. The activity of proteolytic enzymes during diapause has not been investigated. we studied the proteinase activity on four substrates -casein, haemoglobin, bovine serum albumin (bsa), and gelatine -during diapause (from october to march) and in newly hatched males and females in april. during diapause, greater fluctuations in enzyme activity levels were noted in males than in females, and a significant decrease in male enzyme activity was observed in January and march. male enzymes were most effective in decomposing gelatine; whereas, female enzymes were equally effective in hydrolysing gelatine and bsa. The differences in substrate preferences between male and female enzymes were particularly pronounced in october and in the newly hatched individuals. The levels of gelatinolytic activity likely indicate that a high proportion of proteinases in o. rufa are elastase-like enzymes. They are involved in the digestion and remodelling of proteins with numerous peptide bonds formed by amino acids with short side-chains.

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Section: Resultssupporting

confidence: 65%

“…another solitary bee, Megachile rotundata, demonstrated similarly high levels of activity on gelatine (Felicioli et al, 2004). The gelatinolytic activity in O. rufa increased significantly during the late phase of diapause in females and in newly hatched males.…”

Section: Resultsmentioning

confidence: 93%

The Proteolytic Activity of Diapausing and Newly Hatched Red Mason Bees (Osmia Rufa: Megachilidae)

Zaobidna

Dmochowska

Frączek

et al. 2014

Journal of Apicultural Science

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“…The homogenates were then centrifuged at 2500 g for 15 min at 4˚C for debris removal and at 20,000 g for 40 min at the same temperature for supernatant clarification (Felicioli et al, 2004). The insect body extracts from all groups were coded accordingly to the group (as BE, BE-5, and BE-200).…”

Section: Crude Extract Preparations From Beesmentioning

confidence: 99%

“…Extracts of each sample of bees were obtained according to Felicioli et al (2004). Protein concentrations in each sample were determined with the use of the modified BioRad Protein Assay System and the Bradford dye-binding procedure (Bradford, 1976).…”

Section: Crude Extract Preparations From Beesmentioning

confidence: 99%

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The influence of sublethal doses of imidacloprid on protein content and proteolytic activity in honey bees (Apis mellifera L.)

Wilde

Frączek

Siuda

et al. 2016

Journal of Apicultural Research

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The effect of sublethal doses of imidacloprid on protein content and activity of proteases on honey bees was analyzed. The study was conducted in three experimental groups: colonies from groups BE-5 and BE-200 were contaminated with 5 and 200 ppb of imidacloprid, respectively, via their food supply (syrup and pollen), while group BE was used as control (untreated). Bee samples were collected 3 and 10 weeks after feeding started. Protein concentration in bee tissue extracts was analyzed with reference: (a) to the dose of imidacloprid; and (b) duration of exposure to the chemical. The average quantity of protein content was significantly higher at the 3-week interval than in the 10-week interval and the bees from control colonies (BE) had significantly higher protein contents than contaminated bees (BE-5 and BE-200), even 3 weeks after feeding with imidacloprid started. Similarly, the activity of proteolytic enzymes (proteases) was found to be dependent on the dose of imidacloprid used, compared to bees from control colonies showing significantly higher activity.Influencia de las dosis subletales de Imidacloprid en el contenido de proteína y la actividad proteolítica en las abejas melíferas (Apis mellifera L.)Se ha analizado el efecto de las dosis subletales de imidacloprid en el contenido de proteínas y la actividad de las proteasas en las abejas melíferas. El estudio se realizó en tres grupos experimentales: las colonias de los grupos de BE-5 y BE-200 se contaminaron con 5 y 200 ppb de imidacloprid, respectivamente, a través del alimento suministrado (jarabe y polen), mientras que el grupo SER se utilizó como control (sin tratar). Se recogieron muestras de abejas en las semanas 3 y 10 después del comienzo de la alimentació n. La concentració n de proteína en extractos de tejido de abejas se analizó en relació n con: a) la dosis de imidacloprid; y b) la duració n de la exposició n a la sustancia química. La cantidad promedio del contenido de proteína fue significativamente mayor en el intervalo de 3 semanas que en el intervalo de 10 semanas y las abejas de las colonias control (BE) tuvieron un contenido de proteína significativamente más alto que las abejas contaminadas (BE-5 y BE-200), incluso 3 semanas después de que comenzara la alimentació n con imidacloprid. Del mismo modo se encontró que la actividad de las enzimas proteolíticas (proteasas) depende de la dosis de imidacloprid usada, en comparació n con las abejas de colonias de control que muestran una actividad significativamente más alta.

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Proteinase pattern of honeybee prepupae from healthy and American Foulbrood infected bees investigated by zymography

et al. 2018

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American foulbrood disease (AFB) is the main devastating disease that affects honeybees' brood, caused by Paenibacillus larvae. The trend of the research on AFB has addressed the mechanisms by which P. larvae bacteria kill honeybee larvae. Since prepupae could react to the infection of AFB by increasing protease synthesis, the aim of this work was to compare protease activity in worker prepupae belonging to healthy colonies and to colonies affected by AFB. This investigation was performed by zymography. In gel, proteolytic activity was observed in prepupae extracts belonging only to the healthy colonies. In the prepupae extracts, 2D zimography followed by protein identification by MS allowed to detect Trypsin-1 and Chymotrypsin-1, which were not observed in diseased specimens. Further investigations are needed to clarify the involvement of these proteinases in the immune response of honeybee larvae and the mechanisms by which P. larvae inhibits protease production in its host.

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

Cited by 9 publications

References 44 publications

The Proteolytic Activity of Diapausing and Newly Hatched Red Mason Bees (Osmia Rufa: Megachilidae)

The Proteolytic Activity of Diapausing and Newly Hatched Red Mason Bees (Osmia Rufa: Megachilidae)

The influence of sublethal doses of imidacloprid on protein content and proteolytic activity in honey bees (Apis mellifera L.)

Proteinase pattern of honeybee prepupae from healthy and American Foulbrood infected bees investigated by zymography

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