The gene for an abundant parasite coat protein predicts tandemly repetitive metal binding domains.
Immobilization antigens are highly abundant surface membrane proteins that coat the surface of hymenostomatid ciliates. While their function is unknown, recent studies with the common fish parasite, Ichthyophthirius multifiliis, suggest their involvement in a novel mechanism of humoral immunity involving an effect of antibody on parasite behavior. To gain further insight into the nature of these proteins, we have cloned a gene encoding the 48kDa i-antigen of I. multifiliis. Analysis of the gene (designated IAG48[G1]) reveals a single, uninterrupted reading frame that predicts a protein of 442 amino acids. Based on its deduced amino acid sequence, the protein contains hydrophobic amino acid domains at its N- and C-terminus that are characteristic of signal peptide and GPI-anchor addition sites, respectively. The most striking feature of the predicted protein, however, is a series of tandem repeats that spans most of its length. The repeats themselves are characterized by periodic cysteine residues that fall into register when the homologous segments are aligned. Interestingly, the spacing of cysteines (C-X2,3-C) within a framework of larger (C-X2-C-X20-C-X3-C-X20-C-X2-C) motifs is entirely consistent with the structure of known zinc-binding proteins. Finally, comparison of the coding sequence of the 48kDa i-antigen gene with a partial cDNA previously thought to encode this protein reveals nearly complete identity except at their 3' ends, suggesting that alternative forms of the antigen exist.
Clark
T G
TG
Department of Microbiology and Immunology, NYSCVM Cornell University, Ithaca, NY 14853, USA. tgc3@cornell.edu
Lin
T L
TL
Jackwood
D A
DA
Sherrill
J
J
Lin
Y
Y
Dickerson
H W
HW
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology; D000953:Antigens, Protozoan; D000954:Antigens, Surface / Q000235:genetics* / Q000276:immunology; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D005393:Fish Diseases / Q000469:parasitology; D005399:Fishes / Q000276:immunology / Q000469:parasitology*; D017125:Genes, Protozoan; D017261:Glycosylphosphatidylinositols / Q000235:genetics; D016803:Hymenostomatida / Q000235:genetics* / Q000276:immunology / Q000472:pathogenicity; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D015800:Protozoan Proteins / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D020080:Tandem Repeat Sequences / Q000235:genetics
D000906:Antibodies; D000953:Antigens, Protozoan; D000954:Antigens, Surface; D017261:Glycosylphosphatidylinositols; C118810:IAG48 protein, Ichthyophthirius multifiliis; D008565:Membrane Proteins; D015800:Protozoan Proteins; C043323:immobilization antigens