Plasmodium vivax merozoite surface proteins-3beta and-3gamma share structural similarities with P. vivax merozoite surface protein-3alpha 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 within P. vivax lambdagt11 clone 5.4, which was selected by immunoscreening 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 lambdaDASH genomic (g) DNA library. Antibodies to recombinant 5.4 and subsequent fusion proteins produce a pattern of circumferential surface fluorescence by indirect immunofluorescence assays (IFA) on segmented schizonts and free intact merozoites, and recognize a 125 kDa protein via western immunoblots. The gene, however, encodes a protein with a calculated size of 75677 Da, and 3' and 5' RACE analyses were employed to confirm the size of the gene and its coding 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 predicted 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 among the three encoded proteins is low (<40%), the shared predicted structural features and motifs indicate that they are members of an intra-species family, which we are designating as the PvMSP-3 family with the reported members being Pvmsp-3alpha, Pvmsp-3beta, and Pvmsp-3gamma. We further demonstrate that this family also includes related proteins from P. knowlesi and P. falciparum.
Galinski
M R
MR
Emory University, Department of Medicine, Emory Vaccine Research Center, Yerkes Primate Research Center, 954 Gatewood Road, 30329, Atlanta, GA, USA. galinski@rmy.emory.edu
Ingravallo
P
P
Corredor-Medina
C
C
Al-Khedery
B
B
Povoa
M
M
Barnwell
J W
JW
D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan / Q000737:chemistry* / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D001483:Base Sequence; D015153:Blotting, Western; D016054:DNA, Protozoan / Q000032:analysis / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D017125:Genes, Protozoan; D006801:Humans; D016780:Malaria, Vivax / Q000469:parasitology*; D008969:Molecular Sequence Data; D010966:Plasmodium vivax / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D015800:Protozoan Proteins / Q000737:chemistry* / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D012453:Saimiri; D017422:Sequence Analysis, DNA
D000953:Antigens, Protozoan; D016054:DNA, Protozoan; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins; C089133:merozoite surface protein 3, Plasmodium; C436735:merozoite surface protein 3beta; C436736:merozoite surface protein 3gamma