Molecular characterization of a venom acid phosphatase Acph-1-like protein from the Asiatic honeybee Apis cerana

다국어 초록 (Multilingual Abstract)

Bee venom contains a variety of peptides and enzymes, including acid phosphatases. An acid phosphatase hasbeen identified from European honeybee (Apis mellifera) venom. However, although the amino acid sequenceis known, no functional information is currently available for bee venom acid phosphatase Acph-1-like proteins.
In this study, an Asiatic honeybee (Apis cerana) venom acid phosphatase Acph-1-like protein (AcAcph-1) wasidentified. The analysis of the predicted AcAcph-1 amino acid sequence revealed high levels of identity withother bee venom acid phosphatase Acph-1-like proteins. Recombinant AcAcph-1was expressed as a 64-kDa proteinin baculovirus-infected insect cells. The enzymatic properties of recombinant AcAcph-1, determined using pnitrophenylphosphate (p-NPP) as a substrate, showed the highest activity at 45 °C and pH 4.8. Northern andwestern blot analyses showed that AcAcph-1 was expressed in the venom gland and was present as a 64-kDaprotein in bee venom. In addition, N-glycosylation of AcAcph-1was revealed by tunicamycin treatment of recombinantvirus-infected insect Sf9 cells and by glycoprotein staining of purified recombinant AcAcph-1. Our findingsshowthat AcAcph-1 functions as a venomacid phosphatase. This paper provides the first evidence of the role of abee venom acid phosphatase Acph-1-like protein.

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