Plasmodium vivax merozoite surface proteins-3 beta and-3 gamma share structural similarities with P. vivax merozoite surface protein-3 alpha and define a new gene family

The genes encoding two merozoite surface proteins of Plasmodium vivax that are related to PvMSP3 [1] are reported. One of these genes was identified w ithin P. vivax, lambda gt11 clone 5.4, which was selected by immunoscreenin g with a Saimiri monkey antiserum. The insert DNA of this clone was used as a probe to isolate the complete gene from a P. vivax lambda DASH genomic ( g) DNA library. Antibodies to recombinant 5.4 and subsequent fusion protein s produce a pattern of circumferential surface fluorescence by indirect imm unofluorescence assays (IFA) on segmented schizonts and free intact merozoi tes, and recognize a 125 kDa protein via western immunoblots. The gene, how ever, encodes a protein with a calculated size of 75 677 Da, and 3 ' and 5 ' RACE analyses were employed to confirm the size of the gene and its codin g region. The second related P. vivax gene was isolated by hybridization of a fragment of an orthologous P. knowlesi gene. The encoded proteins of all three related P. vivax genes have putative signal peptides, large central domains that contain > 20% alanine residues bound by charged regions, are p redicted to form alpha -helices with heptad repeat coiled-coil structures, and do not have a hydrophobic region that could anchor them to the surface of the merozoite. Although the overall identity in amino acid alignment amo ng the three encoded proteins is low ( < 40%) the shared predicted structur al features and motifs indicate that they are members of an intra-species f amily, which we are designating as the PvMSP-3 family with the reported mem bers being Pvmsp-3 alpha, Pvmsp-3 beta, and Pvmsp-3 gamma. We further demon strate that this family also includes related proteins from P, knowlesi and P. falciparum. (C) 2001 Elsevier Science B.V. All rights reserved.