Characterization of a polymorphic family of integral membrane proteins in promastigotes of different Leishmania species.
Antibodies raised against a Leishmania major recombinant promastigote surface antigen 2 (PSA-2) fragment recognized three major polypeptides of approximate M(r) 96,000, 80,000 and 50,000 in promastigotes of three Israeli isolates of L. major including the cloned line LRC-L137-V121, but detected a different array of polypeptides in other L. major isolates. The pattern was different both in number of polypeptides detected and their molecular weight. The antibodies to L. major PSA-2 also recognized polypeptides in L. tropica, L. donovani and very weakly in L. mexicana promastigotes and in Crithidia lucilliae. The number and size of the polypeptides was different in each species. In addition to the membrane-bound PSA-2 polypeptides we identified water-soluble forms of PSA-2 released in promastigote culture supernatants. Peptide maps of the various L. major PSA-2 membrane polypeptides showed they were different from each other. N-terminal amino acid sequence of the three polypeptides expressed by L. major showed they are similar but distinct, consistent with being members of a polymorphic family. Because of the extensive sequence similarity between the PSA-2 genes it has been difficult to assign protein products to individual genes. As a first step towards solving this problem, we have transfected into L. mexicana a genomic clone of a L. major PSA-2 gene and shown that it produces a M(r) 35,000 polypeptide recognized by monoclonal and polyclonal antibodies to L. major PSA-2.
Walter and Eliza Hall Institute of Medical Research, Post Office Royal Melbourne Hospital, Victoria, Australia.
F M
FM
Symons
P J
PJ
Murray
H
H
Ji
R J
RJ
Simpson
A H
AH
Osborn
R
R
Cappai
E
E
Handman
D000595:Amino Acid Sequence; D000818:Animals; D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan / Q000235:genetics*; D000954:Antigens, Surface / Q000235:genetics*; D015870:Gene Expression; D017125:Genes, Protozoan; D006801:Humans; D007891:Leishmania / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D007893:Leishmania donovani / Q000235:genetics; D018320:Leishmania major / Q000235:genetics; D007894:Leishmania mexicana / Q000235:genetics; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D011110:Polymorphism, Genetic; D015800:Protozoan Proteins / Q000235:genetics* / Q000276:immunology; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity
D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D000954:Antigens, Surface; D015800:Protozoan Proteins; D011994:Recombinant Proteins; C072094:surface antigen P2, Leishmania