The hydrogenosomal enzyme hydrogenase from the anaerobic fungus Neocallimastix sp. L2 is recognized by antibodies, directed against the C-terminal microbody protein targeting signal SKL.
The question was addressed whether antibodies directed against the general microbody C-terminal protein targeting signal SKL recognized hydrogenosomal proteins from Neocallimastix sp. L2. Immunofluorescence, immunocytochemistry and Western blotting experiments using these antibodies indicated the presence of hydrogenosomal proteins containing SKL-COOH. One of these proteins, the hydrogenase, was purified to homogeneity. It has a native molecular mass of 87 kDa and consists of two subunits of approximately 30 and 60 kDa, both cross-reacting with anti-SKL antibodies. Its activity could be inhibited by CO, NO2-, and acetylene, suggesting a (Ni-Fe-Se) hydrogenase. Immunocytochemistry using polyclonal antibodies raised against the hydrogenase revealed the location of this protein in the hydrogenosomal matrix. The results described in this paper suggest that hydrogenosomes from Neocallimastix sp. L2 are related to microbodies from aerobic eukaryotes and support the idea of a common evolutionary origin for these organelles.
Marvin-Sikkema
F D
FD
Department of Microbiology, University of Groningen, Haren/The Netherlands.
Kraak
M N
MN
Veenhuis
M
M
Gottschal
J C
JC
Prins
R A
RA
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D005656:Fungal Proteins / Q000032:analysis* / Q000276:immunology; D005658:Fungi / Q000201:enzymology*; D006864:Hydrogenase / Q000032:analysis*; D007150:Immunohistochemistry; D008830:Microbodies / Q000737:chemistry*; D008969:Molecular Sequence Data; D021382:Protein Sorting Signals / Q000032:analysis*
D000911:Antibodies, Monoclonal; D005656:Fungal Proteins; D021382:Protein Sorting Signals; D006864:Hydrogenase