title: cDNA sequence encoding a Plasmodium falciparum protein associated with knobs and localization of the protein to electron-dense regions in membranes of infected erythrocytes.
abstract: Plasmodium falciparum modifies the host erythrocyte's plasma membrane by the formation of electron-dense structures called knobs. We have produced monoclonal antibodies (McAbs) which specifically bind to the knobs in immunoelectron microscopic experiments with thin sections of parasitized erythrocytes. However, the McAbs fail to bind to the surface of live parasitized erythrocytes. Immunoblotting experiments with these McAbs show the antigen is localized to the erythrocyte plasma membrane. The antigen with which the McAbs react varies in mol. wt from 80 to 95 kd in different knob-producing isolates of P. falciparum and is absent in knobless variants. The McAbs react with the expressed product of a P. falciparum cDNA clone, thus demonstrating that the clone encodes part of this knob-associated protein. The sequence of the cDNA fragment partially overlaps a published cDNA sequence reported to encode the amino-terminal portion of the knob protein, and extends the predicted open reading frame by 190 amino acids. The carboxyl-terminal portion of the predicted amino acid sequence contains a highly charged stretch of approximately 100 amino acid residues. We suggest that this unusual, highly charged region participates in intermolecular salt bridging leading to dense packing of these molecules. This would create the electron-dense regions observed by electron microscopy and might also explain the insolubility of the knob-associated protein in the absence of strong ionic detergents or chaotropic agents.
authors.forename: F
authors.initials: F
authors.lastname: Ardeshir
authors.forename: J E
authors.initials: JE
authors.lastname: Flint
authors.forename: Y
authors.initials: Y
authors.lastname: Matsumoto
authors.forename: M
authors.initials: M
authors.lastname: Aikawa
authors.forename: R T
authors.initials: RT
authors.lastname: Reese
authors.forename: H
authors.initials: H
authors.lastname: Stanley
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000378:metabolism; D004910:Erythrocyte Membrane / Q000469:parasitology* / Q000648:ultrastructure; D006801:Humans; D010455:Peptides / Q000235:genetics*; D010963:Plasmodium falciparum / Q000235:genetics* / Q000648:ultrastructure; D015800:Protozoan Proteins
chemical_list: D010455:Peptides; D015800:Protozoan Proteins; C042968:knob protein, Plasmodium falciparum; D004247:DNA