Antibodies directed against the peroxisomal targeting signal of firefly luciferase recognize multiple mammalian peroxisomal proteins.
We have previously shown that the peroxisomal targeting signal in firefly luciferase consists of the COOH-terminal three amino acids of the protein, serine-lysine-leucine (Gould, S.J., G.A. Keller, N. Hosken, J. Wilkinson, and S. Subramani, 1989. J. Cell Biol. 108:1657-1664). Antibodies were raised against a synthetic peptide that contained this tripeptide at its COOH terminus. Immunofluorescence and immunocryoelectron microscopy revealed that the anti-peptide antibodies specifically detected peroxisomes in mammalian cells. Further characterization revealed that the antibodies were primarily directed against the COOH-terminal three amino acids of the peptide. In Western blot experiments, the antibodies recognized 15-20 rat liver peroxisomal proteins, but reacted with only a few proteins from other subcellular compartments. These results provide independent immunological evidence that the peroxisomal targeting signal identified in firefly luciferase is present in many peroxisomal proteins.
Department of Biology, University of California, La Jolla 92093.
S J
SJ
Gould
S
S
Krisans
G A
GA
Keller
S
S
Subramani
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D001517:Coleoptera / Q000201:enzymology; D000939:Epitopes / Q000032:analysis*; D005455:Fluorescent Antibody Technique; D015248:Gemfibrozil / Q000494:pharmacology; D015151:Immunoblotting; D008099:Liver / Q000187:drug effects / Q000648:ultrastructure; D008156:Luciferases / Q000032:analysis / Q000276:immunology*; D008297:Male; D008830:Microbodies / Q000187:drug effects / Q000648:ultrastructure*; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000276:immunology*; D021382:Protein Sorting Signals / Q000032:analysis / Q000276:immunology*; D011506:Proteins / Q000032:analysis* / Q000276:immunology; D051381:Rats; D011919:Rats, Inbred Strains; D012016:Reference Values
D000939:Epitopes; D009842:Oligopeptides; D021382:Protein Sorting Signals; D011506:Proteins; D008156:Luciferases; D015248:Gemfibrozil