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Identification of the proteome complement of high royal jelly producing bees (Apis mellifera) during worker larval development

Analyse du protéome, lors du développement larvaire, des ouvrières d’une lignée d’abeilles (Apis mellifera) produisant de grandes quantités de gelée royale

Analyse des Proteoms der Larvalentwicklung von Arbeiterinnen einer Zuchtlinien der Honigbiene (Apis mellifera), die in hohen Mengen Gelée Royale produziert

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Abstract

To investigate the composition and function of the proteome during larval development of worker bees from a strain of Apis mellifera L. artificially selected for increased royal jelly yield, proteins were partially identified by two-dimensional gel electrophoresis, mass spectrometry and protein engine identification tools that were applied to the honeybee genome. Out of 48 high abundance proteins selected for MS fingerprinting, 22 could be identified as representing annotated genes of the honey bee. These including 9 nutrient related proteins, 6 proteins associated with carbohydrate metabolism and energy production, 3 heat shock proteins, 4 other proteins related to the metabolism of amino acids, fatty acid metabolism, larval growth and cell cycle, respectively.

Zusammenfassung

Mit der Verfügbarkeit des komplett sequenzierten und annotierten Genoms der Honigbiene liegen jetzt Proteomanalysen im Rahmen der Möglichkeit. Obwohl einzelne Gene, Proteine und Enzyme des Metabolismus bereits seit einiger Zeit intensiv untersucht werden, stecken generelle Proteomanalysen, z.B. zum Proteom der Larvalentwicklung, noch in den Kinderschuhen. Ziel dieser Studie war eine systematische Untersuchung der Proteine, die in diesem Entwicklungsprozess eine Rolle spielen. Der experimentelle Ansatz bestand in einer zweidimensionalen elektrophoretischen Auftrennung gefolgt von MALDI-TOF Analysen der einzelnen Proteinspots.

Aus der Gesamtzahl der Proteine, die in der Larvalentwicklung zu finden waren, identifizierten wir 22 Spots (Abb. 1; Tab. I). Diese umfassten 9 Nahrungsproteine, 6 Proteine mit Bezug zum Kohlenhydratstoffwechsel und zur Energieproduktion, 3 Hitzeschockproteine, sowie 4 weitere Proteine mit Bezug zum Aminosäurenstoffwechsel, Fettsäurenstoffwechsel, Zellzyklus und zur Larvalentwicklung allgemein. Einige der hier identifizierten Proteine (z.B. die Gelée Royal Proteine) stammen vermutlich aus dem Darm und stellen so einen Teil der Larvennahrung dar, aber nicht des larvalen Proteoms im eigentlichen Sinne.

Das Larvale Serumprotein 2 findet sich vor allem an Tag 6 der Entwicklung stark angereichert und könnte eine Rolle in der Speicherung von Aminosäuren für die Metamorphose spielen (Abb. 1 und 2). Vier der im Kohlenhydratstoffwechsel und der Energieproduktion wichtigen Proteine, Enolase, Argininkinase, Aldehyddehydrogenase und Phosphoglyzeromutase zeigten eine ansteigende Expression in der Entwicklung (Abb. 1 und 2). ATP-Synthase zeigte ein unverändertes Expressionsmuster, während der Vorläufer einer alpha-Kette der mitochondrialen ATP-Synthase zunehmend weniger exprimiert wird. Diese Befunden deuten darauf hin, dass die Entwicklung energieabhängig ist. Die Persistenz von Hitzeschockproteinen im larvalen Proteom (des mitochondrialen Vorläufers eines 60 kDa Hsp, des Hitzeschock-ähnlichen Proteins 3 und der Isoform 1 des Hitzeschockproteins 8) kann mit ihrer Rolle in der Proteinfaltung erklärt werden (Abb. 1 und 2). Das Protein Lethal (2) 37Cc spielt in der DNA-Reparatur und im Zellzyklus eine Rolle und ist hier vermutlich für den larvalen Stoffwechsel und den Übergang zur Puppenphase von Bedeutung. Der mit dem Aminosäuren- und Nitratstoffwechsel in Verbindung stehende Vorläufer des Enzyms Ornithin-Aminotransferase und das im Fettstofwechsel wichtige Fettsäuren-Bindungsprotein zeigten beide einen Anstieg der Expression im Verlauf der Larvalentwicklung (Abb. 1 und 2).

Diese Ergebnisse zeigen einerseits, dass die Larvalentwicklung ein dynamischer Prozess ist, der eine Vielzahl an Proteinen mit unterschiedlichen Funktionen erfordert. Andererseits sind dies jedoch nur vorläufig Ergebnisse, da nur ein kleiner Teil der Proteinspots tatsächlich identifiziert werden konnte, und somit weitere Untersuchungen erforderlich sind.

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Authors and Affiliations

  1. Institute of Apicultural Research, Chinese Academy of Agricultural Science, 100093, Beijing, China

    Jianke Li & Huawei Li

  2. Bioengineering Department, Zhengzhou University, 450001, Zhengzhou, China

    Huawei Li

  3. Institute of Crop Science, Chinese Academy of Agricultural Sciences, The National Key Facility for Crop Gene Resources and Genetic Improvement, 12 Zhongguancun Nandajie, 100081, Beijing, China

    Zhaohui Zhang & Yinghong Pan

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  1. Jianke Li
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  2. Huawei Li
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Correspondence to Yinghong Pan.

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Manuscript editor: Marla Spivak

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Li, J., Li, H., Zhang, Z. et al. Identification of the proteome complement of high royal jelly producing bees (Apis mellifera) during worker larval development. Apidologie 38, 545–557 (2007). https://doi.org/10.1051/apido:2007047

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