Profile of the body surface proteolytic system in Apis mellifera queens
2011
Strachecka, A.J.,University of Life Sciences, Lublin (Poland). Dept. of Biological Basis of Animal Production | Gryzinska, M.M.,University of Life Sciences, Lublin (Poland). Dept. of Biological Basis of Animal Production | Krauze, M.,University of Life Sciences, Lublin (Poland). Dept. of Biochemistry and Toxicology | Grzywnowicz, K.,Maria Curie-Sklodowska Univ., Lublin (Poland). Dept. of Biochemistry
The aim of this study was to examine proteolytic activity on the body surface of queen honey bees at their various developmental stages (eggs, larvae, pupae and imagines) and in all four seasons. Extracts of the body surface material with water and detergent were used for an in vitro analysis of the proteolytic activity and protease inhibitor level, as well as for an electrophoretic separation of the extracts in polyacrylamide gels. The following methods were used: protein content tested by the Lowry method (modified by Schacterle-Pollack), protease activity tested by the Anson method and protease inhibitor activity tested by the Lee and Lin method. Our studies revealed a high protease activity in an acidic environment (pH = 2.4; the material rinsed with detergent), as well as in neutral (pH = 7) and alkaline (pH = 11.2) environments (the material rinsed with water in both cases). The highest protein concentration values were observed in the imagines from summer. The lowest activities of the proteases and protease inhibitors were determined in the eggs from summer. The highest activities of the acidic, neutral and alkaline proteases were observed in the pupae from spring. The highest number of protease activity bands in PAGE zymography was obtained for the neutral and alkaline activities in the queens in all seasons. In the queens all the catalytic protease types were present: asparagine and cysteine proteases at pH = 2.4; cysteine proteases and metalloproteases at pH = 7 and serine proteases at pH = 11.2. These results were crucial for the analysis of immunity mechanisms on the body surface of the honey bee.
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