Molecular characterization of a venom acid phosphatase Acph-1-like protein from the Asiatic honeybee Apis cerana |
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Institution: | 1. Honeybee Research Institute, Jiangxi Agricultural University, Nanchang 330045, PR China;2. Department of Biological Sciences, Macquarie University, Building W19F, North Ryde, NSW 2109, Australia;1. Department of Plant Protection, College of Agriculture Sciences, University of Guilan, Rasht, Iran;2. Department of Plant Protection, Faculty of Agriculture and Natural Resources, Higher Education Complex of Shirvan, Iran;3. Biocontrol and Insect Pathology Laboratory, Department of Plant Protection, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran |
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Abstract: | Bee venom contains a variety of peptides and enzymes, including acid phosphatases. An acid phosphatase has been identified from European honeybee (Apis mellifera) venom. However, although the amino acid sequence is 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) was identified. The analysis of the predicted AcAcph-1 amino acid sequence revealed high levels of identity with other bee venom acid phosphatase Acph-1-like proteins. Recombinant AcAcph-1 was expressed as a 64-kDa protein in baculovirus-infected insect cells. The enzymatic properties of recombinant AcAcph-1, determined using p-nitrophenyl phosphate (p-NPP) as a substrate, showed the highest activity at 45 °C and pH 4.8. Northern and western blot analyses showed that AcAcph-1 was expressed in the venom gland and was present as a 64-kDa protein in bee venom. In addition, N-glycosylation of AcAcph-1 was revealed by tunicamycin treatment of recombinant virus-infected insect Sf9 cells and by glycoprotein staining of purified recombinant AcAcph-1. Our findings show that AcAcph-1 functions as a venom acid phosphatase. This paper provides the first evidence of the role of a bee venom acid phosphatase Acph-1-like protein. |
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