Antibodies directed against the peroxisomal targeting signal of firefly luciferase recognize multiple mammalian peroxisomal proteins

Stephen J Gould, Skaidrite Krisans, Gilbert Andre Keller, Suresh Subramani

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish (US)
Pages (from-to)27-34
Number of pages8
JournalJournal of Cell Biology
Volume110
Issue number1
StatePublished - Jan 1990
Externally publishedYes

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Firefly Luciferases
Antibodies
Peptides
Proteins
Amino Acids
Peroxisomes
Fluorescence Microscopy
Leucine
Serine
Lysine
Anti-Idiotypic Antibodies
Western Blotting
Liver

ASJC Scopus subject areas

  • Cell Biology

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Antibodies directed against the peroxisomal targeting signal of firefly luciferase recognize multiple mammalian peroxisomal proteins. / Gould, Stephen J; Krisans, Skaidrite; Keller, Gilbert Andre; Subramani, Suresh.

In: Journal of Cell Biology, Vol. 110, No. 1, 01.1990, p. 27-34.

Research output: Contribution to journalArticle

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