Import of human bifunctional enzyme into peroxisomes of human hepatoma cells in vitro.
A polypeptide containing the carboxyl-terminal fragment of human peroxisomal enoyl-CoA hydratase:3-hydroxyacyl-CoA dehydrogenase bifunctional enzyme was synthesized in vitro from its cDNA clone. This expression polypeptide was transported into purified rat liver peroxisomes. When the expression polypeptide was incubated with postnuclear supernatant fractions of human hepatoma cells and analyzed by Nycodenz gradient SDS-PAGE and fluorography, it was imported specifically into peroxisomes as indicated by its resistance to proteinase K degradation. A deletion of the last nine amino acid residues at the carboxyl-terminus of this polypeptide prevents its peroxisomal import. A tripeptide sequence, SKL, located at the carboxyl-terminus of human bifunctional enzyme appears to be the targeting signal for the peroxisomal importation of bifunctional enzyme in human cells.
Chen
G L
GL
Kennedy Institute, School of Medicine, Johns Hopkins University, Baltimore, MD 21205.
Balfe
A
A
Erwa
W
W
Hoefler
G
G
Gaertner
J
J
Aikawa
J
J
Chen
W W
WW
D015094:3-Hydroxyacyl CoA Dehydrogenases / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D006528:Carcinoma, Hepatocellular / Q000201:enzymology* / Q000235:genetics; D002460:Cell Line; D003001:Cloning, Molecular; D004746:Enoyl-CoA Hydratase / Q000235:genetics*; D006801:Humans; D007535:Isomerases / Q000235:genetics*; D008099:Liver / Q000201:enzymology; D008113:Liver Neoplasms / Q000201:enzymology* / Q000235:genetics; D008830:Microbodies / Q000201:enzymology*; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000235:genetics*; D063994:Peroxisomal Bifunctional Enzyme; D014176:Protein Biosynthesis; D051381:Rats; D012689:Sequence Homology, Nucleic Acid; D014158:Transcription, Genetic
D009097:Multienzyme Complexes; D015094:3-Hydroxyacyl CoA Dehydrogenases; C576253:EHHADH protein, human; D004746:Enoyl-CoA Hydratase; D063994:Peroxisomal Bifunctional Enzyme; D007535:Isomerases