The merozoite surface protein 6 gene codes for a 36 kDa protein associatedwith the Plasmodium falciparum merozoite surface protein-1 complex

A complex of non-covalently bound polypeptides is located on the surface of the merozoite form of the human malaria parasite Plasmodium falciparum. Fo ur of these polypeptides are derived by proteolytic processing of the meroz oite surface protein 1 (MSP-1) precursor. Two components, a 22 and a 36 kDa polypeptide are not derived from MSP-1. The N-terminal sequence of the 36 kDa polypeptide has been determined, the corresponding gene cloned, and the protein characterised. The 36 kDa protein consists of 211 amino acids and is derived from a larger precursor of 371 amino acids. The precursor merozo ite surface protein 6 (MSP-6) has been designated, and the 36 kDa protein, MSP-6(36). Mass spectrometric analysis of peptides released from the polype ptide by tryptic digestion confirmed that the gene identified codes for MSP -6(36). Antibodies were produced to a recombinant protein containing the C- terminal 45 amino acid residues of MSP-6(36). In immunofluorescence studies these antibodies bound to antigen at the parasite surface or in the parasi tophorous vacuole within schizonts, with a pattern indistinguishable from t hat of antibodies to MSP-1. MSP-6(36) was present in the MSP-1 complex immu noprecipitated from the supernatant of in vitro parasite cultures, but was also immunoprecipitated from this supernatant in a form not bound to MSP-1. Examination of the MSP-6 gene in three parasite lines detected no sequence variation. The sequence of MSP-6(36) is related to that of the previously described merozoite surface protein 3 (MSP-3). The MSP-6(36) amino acid seq uence has 50% identity and 85% similarity with the C-terminal region of MSP -3. The proteins share a specific sequence pattern (ILGWEFGGG-[AV]-P) and a glutamic acid-rich region. The remainder of MSP-6 and MSP-3 are unrelated, except at the N-terminus. Both MSP-6(36) and MSP-3 are partially associate d with the parasite surface and partially released as soluble proteins on m erozoite release. MSP-6(36) is a hydrophilic negatively charged polypeptide , but there are two clusters of hydrophobic amino acids at the C-terminus, located in two amphipathic helical structures identified from secondary str ucture predictions. It was suggested that this 35 residue C-terminal region may be involved in MSP-6(36) binding to MSP-1 or other molecules; alternat ively, based on the secondary structure and coil formation predictions, the region may form an intramolecular anti-parallel coiled-coil structure. (C) 2001 Elsevier Science B.V. All rights reserved.