A novel 40-kDa membrane-associated EF-hand calcium-binding protein in Plasmodium falciparum.
A 40-kDa sexual stage radiolabeled surface protein of Plasmodium falciparum, Pfs40, was previously identified as a potential target antigen of transmission blocking immunity by an immunogenetic approach. Synthetic oligonucleotide "guessmers," based on microsequenced tryptic peptides of Pfs40 purified by two-dimensional gel electrophoresis, were used to clone the full length cDNA and genomic DNA encoding Pfs40. The deduced amino acid sequence predicted an integral membrane protein containing five EF-hand calcium-binding domains. The biological activity of one or more of these domains was confirmed by binding of 45Ca to both native and recombinant Pfs40. Antisera to recombinant Pfs40 immunoprecipitated the native radiolabeled 40-kDa surface protein. The predicted noncytosolic membrane-associated localization of Pfs40 is unique within the EF-hand calcium-binding protein superfamily.
Rawlings
D J
DJ
Malaria Section, National Institutes of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892.
Kaslow
D C
DC
D000595:Amino Acid Sequence; D000818:Animals; D001345:Autoradiography; D001483:Base Sequence; D001665:Binding Sites; D015153:Blotting, Western; D002135:Calcium-Binding Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D003001:Cloning, Molecular; D016054:DNA, Protozoan / Q000235:genetics; D016521:Electrophoresis, Gel, Pulsed-Field; D004591:Electrophoresis, Polyacrylamide Gel; D008565:Membrane Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D008969:Molecular Sequence Data; D010963:Plasmodium falciparum / Q000235:genetics / Q000378:metabolism*; D011233:Precipitin Tests; D015800:Protozoan Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism
D002135:Calcium-Binding Proteins; D016054:DNA, Protozoan; D008565:Membrane Proteins; D015800:Protozoan Proteins; C073035:Pf39 protein, Plasmodium falciparum