Use of phage display technology to identify antigenic sites in major royal jelly proteins and localization of MRJPs putative binding proteins in the honeybee brain.
Leonardo Gomes Peixoto; Fausto Emilio Capparelli; Luciana Karen Calábria; Claudia Tavares dos Santos; Ana Clara Silveira Broggine; Luis Ricardo Goulart Filho; Foued Salmen Espindola.
Instituto de Genética e Bioquímica, Universidade Federal de Uberlândia.
The major royal jelly proteins (MRJPs) is a unique protein family highly expressed in the hypopharyngeal gland of worker honeybee (Apis mellifera) that are related at the sequence level to the yellow protein family in Drosophila. MRJP1 or apalbumin-1 is the major bee-milk component used to feed the queen larva. Other study showed in the bee brain the MRJP1 mRNA expression. We detected in the bee brain homogenate three polypeptides, p57, p70 and p128, that cross-reacted with an antibody to MRJP1. In this study, we use this antibody and phage display approach to identify MRJP1 antigenic sites and localize its putative binding proteins in the brain. In this way, 21 peptides of the positive phage homogenate were selected by biopanning based in the reactivity to MRJP1 antibody. Mapping these peptides sequences as binding sites to anti-MRJP1 we verified that its showed very similar sequences to all the others MRJPs available in the data bank. This peptides pool were immunolocalized as putative binding partners in honeybee brain areas such as the mushroom bodies and optic lobes as well as in the hypopharyngeal gland to the worker, drone and queen. This study suggested that putative MRJPs binding proteins may have specific brain and gland distribution upon caste and behavioral features.
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