Abstract
We investigated the protein complement of the hypopharyngeal gland (HG) of winter worker bees from a strain of Apis mellifera artificially selected for increased royal jelly yield and A. m. carnica winter worker bees. Proteins were partially identified using two-dimensional gel electrophoresis (2-DE). MALDI-TOF MS and protein engine identification tools that were utilized for the honeybee genome. Most identified proteins in the two bees strains were assigned to major royal jelly (RJ) proteins (MRJPs). Marked differences were found in the heterogeneity of the MRJPs, in particular MRJP3. Two of the proteins, α-glucosidase and glucose oxidase, were related to carbohydrate metabolism and energy. For the first time in the HG of honeybees, two proteins, peroxiredoxin and thioredoxin peroxidase, which are related to antioxidation functions, and actin 5C, a major cytoskeletal actin protein which may supply enough actin for normal function of cells, have been identified. Results suggest that the HGs serve a storage function in winter and that during the winter period the HG of high RJ producing bees store more proteins than those of Carnica bees.
Zusammenfassung
Mit der Verfügbarkeit eines komplett sequenzierten Genoms ergeben sich jetzt neue Einsichten sowohl für die Honigbiene selbst, als auch für Vergleiche mit anderen Arten. Obwohl einzelne Gene, Proteine und Enzyme des Metabolismus bereits seit einiger Zeit intensiv untersucht werden, stecken generelle Proteomanalysen, z.B. zum Proteom der Hypopharynxdrüsen, noch in den Anfängen. Ziel dieser Studie war eine systematische Untersuchung der Proteinzusammensetzung der Hypopharynxdrüse (HD) von Winterbienen. Der experimentelle Ansatz bestand in einer zweidimensionalen elektrophoretischen Auftrennung gefolgt von MALDI-TOF Analysen der einzelnen Proteinspots.
Aus der Gesamtzahl der Proteine, die in den HD von Winterbienen zu finden waren, identifizierten wir 26 Spots für die HD von Carnica-Arbeiterinnen (Abb. 1A; Tab. I) und 34 Spots für die HD einer chinesischen Zuchtlinie, die in hohen Mengen Gelée royale produziert (viel-Gelée-royal-Linie) (Abb. 1B; Tab. II). Die meisten dieser Proteine (20 für Carnica und 34 für die viel-Gelée-royal-Linie) waren Gelée royal Proteine (GRPs) und dies überwiegend Isoformen von GRP 3. Mit Bezug zum Kohlenhydratstoffwechsel fanden wir drei Proteine, zwei Glucoseoxidasen und eine alpha-Glucosidase. Drei Proteine wurden in dieser Studie zum ersten Mal für Honigbienen beschrieben. Zwei dieser Proteine, Peroxiredoxin und Thioredoxinperoxidase haben eine antioxidante Funktion. Das dritte Protein war ein Actin, die Isoform 1 von Act5C. Die Gesamtzahl der für die viel-Gelée-royal-Linie identifizierten Proteine lag deutlich über der für Carnica Winterbienen, vor allem was die GRPs angeht. Keine Unterschiede zwischen den beiden Linien fanden wir für drei der Kohlenhydratstoffwechselproteine und die drei neubeschriebenen Proteine (Abb. 2B). Diese vorläufigen Ergebnisse zum Proteom der Hypoharynxdrüse deuten darauf hin, dass diese Drüse bei Winterbienen eine Speicherfunktion haben könnte. Ausserdem konnten wir zeigen, dass die HD der viel-Gelée-royal-Linie mehr Proteine enthält als die der Carnica Arbeiterinnen. Da diese Studie nur einen Teil der Proteine des Gesamproteoms der HD identifizieren konnte, sind weitere Untersuchungen erforderlich.
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Li, J., Feng, M., Zhang, Z. et al. Identification of the proteome complement of hypopharyngeal glands from two strains of honeybees (Apis mellifera). Apidologie 39, 199–214 (2008). https://doi.org/10.1051/apido:2007059
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DOI: https://doi.org/10.1051/apido:2007059