| Royal jelly, secretion of hypopharyngeal glands of nurse honeybee wokers, is anatural health promotion bee-products, which makes important contributions toimprove the level of human health. To date, China has selected a honeybee strain,named jelly bees (Apis mellifera L.), that significantly increased royal jellyproduction and it is aslo the imporpant honeybee resource of our country. Dispectresearch efferts have been made to reveal the mechanism of increasing royal jellyoutputs at level of cytology, genetics and molecule, understanding the molecularmechanism of enhanced royal jelly yields at phosphoproteomics level still remianunclear yet.Phosphopeptides of hypopharyngeal gland at newly emerged, nurse and foragerworker bees were enriched by IMAC (immobilized metal-affinity chromatography)and TiO2(titanium dioxide) chromatography, followed by LC-MS (liquidchromatography-mass spectromitry) identification and bioinformatics analysis. Of the971phosphoproteins identified from three aged hypopharyngeal gland,553,734,710,phosphoproteins were identified in newly emerged, nurse and forager bees,respectively. All these proteins were mainly involed in61biological pathways, suchas metabolic pathway of amino acid, nucleotide and fatty acid and CDK2, CDK3,CK2major kinase families.The hypopharyngeal glands requires a core phosphoproteome (387phosphoproteins commonly presented by three aged glands,39.7%of the971phosphoproteins) to regulate its development. The phosphoproteins in the corephosphoproteome were mainly implicated in protein synthesis by intensivelyinvolment of transcriptional and translational mechinary, folding and transport of newsynthesed proteins. They were mainly participated in the pathways of amino acids,nucleotides and energy metabolism, and their phosphorylation were induced byCDK2、CDK3、PKC kinase families. All these phosphorylated proteins and inducedpathways and kiniese are supposed to be a driving force to promote cell proliferation, provide metabolic energy and tissue building block as proteins and nucleotides for thedeveloping glands.The biological functions of hypopharyngeal gland at different developmentalstages are quite different. This has reflected in our data that specificphosphoproteome, pathways and kinase families were indentifed to match with thesefunctional disparaceny of hypopharyngeal glands developing at different stages. Thehypopharyngeal glands of newly emerged bees still young and need rapiddevelopment to get a physioligcal ready for royal jelly secretion in the up-comingnurse stage. Towards this goal, phosphoproteins related to the mechinary oftranscription, translation and transport were highly expressed. Meanwhile the strongeractivated pathways of energy metabolism and CDK2、CDK3、PKA kinase are belivedto promote cell proliferation and satisfy the needs of proteins and energy for fastgrowing glands.The main roles of hypopharyngeal glands of nurse bees are the secretion ofprotein-riched royal jelly, this thus demand the glands synthese large bulk of proteinsat this stage. To this effect, the stronger expressed phosphoproteins related totranscriptional and translational mechinary, and iduced pathways of protein synthesis,CK2and PKA kinases signify their key roles in promoting cell proliferation,enhancing the capacity protein syethesis of royal jelly. The high level expression ofphosphorylated major royal jelly proteins is likely to enhance the nutrition efficiencyfor honeybees via keep the balance of calcium and phosphate in royal jelly that isvital important for fast growing larva. To timely secret royal jelly to the young larvae,the brood pheromone must stimulate the nurse bees and attract them to feed them, thisis achived by highly expressed signal transductional phosphoproteins.The functionality of hypopharyngeal gland of forager has thansformed fromroyal jelly seceretion to the secreting enzyme converting nectar into honey. Thisfunctional changes of the glands require remodeling of cell. To this end, highlyexpressed phosphoproteins related to transcriptional and translational mechinary, andtheir higly implicated in the pathway of energy metabolism and CDK2, CDK3andPKC kinase may serve these purpose. The most striking feature at this stage is thatphosphoproteins were intensive participated in nucleotide metabolism pathways. The metabolic products in nucleotide metabolism pathways involved in signaltransduction, which promote the glands to synthesis and secretion of nectar invertasein accordance with the internal and external needs of pollen of colony. |
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