Presence and cellular distribution of a 60-kDa protein related to mitochondrial hsp60 in Giardia lamblia.
Giardia lamblia trophozoites contain a 60-kDa protein recognized in immunoblots by antibody to mammalian hsp60, a protein localized in mitochondria in eukaryotic cells. The cellular distribution of this protein is evaluated by immunofluorescence microscopy using monoclonal antibody to human hsp60, polyclonal antibody to rodent hsp60, and 2 monoclonal antibodies to mycobacterial 65-kDa antigen, a prokaryotic hsp60 homolog. All of these antibodies, except 1, which is specific for prokaryotic hsp60, give a punctate labeling pattern throughout the cytoplasm, indicating that the 60-kDa protein is concentrated at discrete sites in the cytoplasm. The polyclonal hsp60 antiserum reveals additional punctate labeling colocalized on axonemes of the anterior flagella. Postembedment immunogold labeling and electron microscopy confirm that the antigen is clustered at foci in the cytoplasm but show no evidence of association with a membranous organelle. The hsp60 reactivity is also observed on anterior axonemes within the cytoplasm and on the adhesive disc. Hoechst 33258 DNA staining as well as electron microscopy give no evidence of endosymbionts, so a false positive due to a prokaryotic hsp60 homolog is unlikely. The presence of an hsp60-related protein in G. lamblia raises interesting questions concerning its origin.
Soltys
B J
BJ
Department of Biochemistry, McMaster University, Hamilton, Ontario, Canada.
Gupta
R S
RS
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000913:Antibodies, Protozoan / Q000276:immunology; D015153:Blotting, Western; D018834:Chaperonin 60; D003429:Cross Reactions; D016054:DNA, Protozoan / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D016829:Giardia lamblia / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure; D006360:Heat-Shock Proteins / Q000032:analysis* / Q000276:immunology; D016253:Microscopy, Immunoelectron; D015800:Protozoan Proteins / Q000032:analysis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D000913:Antibodies, Protozoan; D018834:Chaperonin 60; D016054:DNA, Protozoan; D006360:Heat-Shock Proteins; D015800:Protozoan Proteins