Procyclin: an unusual immunodominant glycoprotein surface antigen from the procyclic stage of African trypanosomes.
An immunodominant species-specific surface glycoprotein antigen was purified from procyclic culture forms of Trypanosoma brucei rhodesiense using lectin affinity chromatography and a monoclonal antibody immunoadsorbent. The purified molecule appears on a 10% polyacrylamide gel as a wide, dark silver staining band having an apparent molecular mass of between 30 and 40 kDa, identical to that revealed by immunoblotting using anti-procyclic lysates. The molecule, which we have named procyclin, was shown by immunofluorescence and immunoelectron microscopy to be exposed on the surface of procyclic trypanosomes. Gas-phase protein microsequencing and micro-amino acid analysis revealed an unusual acidic polypeptide with an amino-terminal amino acid sequence which matched portions of previously published sequences predicted from two different cDNAs obtained using mRNA from procyclic trypanosomes. The procyclin molecules contained a large glutamic acid-proline repeat and the form we isolated was highly water soluble. Ten different monoclonal antibodies were used in ELISA with synthetic peptides to localize parasite surface epitopes to various portions of procyclin. The results showed that surface epitopes were spread throughout most of the procyclin molecule, including the glutamic acid-proline repeat portion. Procyclin is distributed over the surface of both culture form and tsetse fly midgut form procyclic trypanosomes, is developmentally regulated and is immunologically species-specific.
Richardson
J P
JP
Department of Biochemistry and Microbiology, University of Victoria, B.C., Canada.
Beecroft
R P
RP
Tolson
D L
DL
Liu
M K
MK
Pearson
T W
TW
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000953:Antigens, Protozoan / Q000302:isolation & purification*; D000954:Antigens, Surface / Q000302:isolation & purification*; D000939:Epitopes / Q000032:analysis; D007118:Immunoassay; D011817:Rabbits; D014346:Trypanosoma brucei brucei / Q000276:immunology*; D014643:Variant Surface Glycoproteins, Trypanosoma / Q000302:isolation & purification*
D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan; D000954:Antigens, Surface; D000939:Epitopes; D014643:Variant Surface Glycoproteins, Trypanosoma