Biochemical characterization of the protective membrane glycoprotein GP46/M-2 of Leishmania amazonensis

Biochemical features of the immunologically protective, membrane glycoprotein GP46/M-2 of Leishmania amazonensis have been investigated. The protein appears to have a single carbohydrate side chain of approximately 3 kDa, representing 7% of the mass of the mature GP46/M-2 protein. Experiments removing this carbohydrate side chain from GP46/M-2 indicate that the carbohydrate is not involved in the epitope recognized by the monoclonal antibody, M-2. As this monoclonal antibody recognizes a species-specific epitope, these data suggest that this determinant is defined by the polypeptide portion of the molecule. Studies employing the VSG-lipase as well as anti-CRD antibody clearly indicate that the molecule is anchored to the surface membrane of the promastigote via a phosphatidylinositol-linked lipid anchor. Neither the carbohydrate side chain nor the lipid anchor appear to be responsible for the apparent refractoriness of this protein to protease digestion, suggesting that properties of the polypeptide itself may be responsible. These data are discussed in terms of recent DNA-derived protein sequence of the GP46/M-2.