Zymolyase treatment exposes p55 antigen of Pneumocystis carinii.
Rats exposed to Pneumocystis carinii mount antibody responses to a broad band migrating on western blot with an apparent molecular weight of 45-55 kDa. One antigen within this band, designated p55, is uniformly recognized by P. carinii exposed rats. Although the gene encoding the p55 antigen had been previously cloned, the location of this antigen within the organism was unknown. Prior attempts to localize the protein were unsuccessful. A monospecific polyclonal antiserum raised against a carboxyl-terminal 15-oligomer peptide yielded specific reactivity with a single 55 kDa band on a western blot of P. carinii. Using this antiserum, little to no reactivity could be detected with P. carinii organisms by immunofluorescence assay (IFA). However, zymolyase treatment of P. carinii dramatically increased the intensity and proportion of organisms reactive by IFA. Zymolyase, an enzyme with beta-1,3 glucanase activity, has previously been shown to remove the electron dense outer layer of the P. carinii cell wall, exposing an electron lucent layer. Immunoelectron microscopy performed on zymolyase treated organisms showed the majority of labeling occurs within the cell wall.
Broomall
K R
KR
Department of Cell Biology, Neurobiology, and Anatomy, University of Cincinnati College of Medicine, Ohio, USA.
Morris
R E
RE
Walzer
P D
PD
Smulian
A G
AG
D000818:Animals; D000913:Antibodies, Protozoan / Q000276:immunology; D000946:Antigens, Fungal / Q000032:analysis*; D015153:Blotting, Western; D002473:Cell Wall / Q000276:immunology; D005455:Fluorescent Antibody Technique; D005656:Fungal Proteins / Q000032:analysis*; D006867:Hydrolases / Q000378:metabolism; D016253:Microscopy, Immunoelectron; D011010:Pneumocystis / Q000276:immunology* / Q000302:isolation & purification; D016720:Pneumocystis Infections / Q000276:immunology; D051381:Rats; D011917:Rats, Inbred Lew
C080689:55 kDa protein, Pneumocystis carinii; D000913:Antibodies, Protozoan; D000946:Antigens, Fungal; D005656:Fungal Proteins; C037833:zymolyase; D006867:Hydrolases