Biochemical isolation of an insect haemocyte anti-aggregation protein from the venom of the endoparasitic wasp, Pimpla hypochondriaca, and identification of its gene

Pimpla hypochondriaca venom is complex and contains a number of different proteins and polypeptides that exert a variety of effects on insect physiology. In particular, it possesses factors with potent anti-haemocyte and immunosuppressive properties. In the current work, we describe the biochemical isolation of a single venom factor with insect haemocyte anti-aggregation properties. The protein was isolated using gel filtration and ion exchange chromatography, in conjunction with a qualitative in vitro haemocyte anti-aggregation assay to monitor activity and confirm identity. The protein has a molecular weight estimate of 33 kDa (determined by SDS PAGE under reducing conditions), and an N-terminal sequence of Asp-Ser-Asp-Ile-Tyr-Leu-Leu. The biochemically isolated protein has been demonstrated to inhibit haemocyte aggregation and to suppress encapsulation responses, using in vitro and in vivo assays, respectively. Furthermore, its gene has been identified as vpr3. The work is presented within the context of the role of P. hypochondriaca venom and the isolated protein in host immune suppression.