title: Peroxisome targeting signal of rat liver acyl-coenzyme A oxidase resides at the carboxy terminus.
abstract: To identify the topogenic signal of peroxisomal acyl-coenzyme A oxidase (AOX) of rat liver, we carried out in vitro import experiments with mutant polypeptides of the enzyme. Full-length AOX and polypeptides that were truncated at the N-terminal region were efficiently imported into peroxisomes, as determined by resistance to externally added proteinase K. Polypeptides carrying internal deletions in the C-terminal region exhibited much lower import activities. Polypeptides that were truncated or mutated at the extreme C terminus were totally import negative. When the five amino acid residues at the extreme C terminus were attached to some of the import-negative polypeptides, the import activities were rescued. Moreover, the C-terminal 199 and 70 amino acid residues of AOX directed fusion proteins with two bacterial enzymes to peroxisomes. These results are interpreted to mean that the peroxisome targeting signal of AOX residues at the C terminus and the five or fewer residues at the extreme terminus have an obligatory function in targeting. The C-terminal internal region also has an important role for efficient import, possibly through a conformational effect.
authors.affiliation: Department of Biochemistry, Shinshu University School of Medicine, Nagano, Japan.
authors.forename: S
authors.initials: S
authors.lastname: Miyazawa
authors.forename: T
authors.initials: T
authors.lastname: Osumi
authors.forename: T
authors.initials: T
authors.lastname: Hashimoto
authors.forename: K
authors.initials: K
authors.lastname: Ohno
authors.forename: S
authors.initials: S
authors.lastname: Miura
authors.forename: Y
authors.initials: Y
authors.lastname: Fujiki
mesh_terms: D000214:Acyl Coenzyme A / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis; D008099:Liver / Q000201:enzymology*; D008830:Microbodies / Q000201:enzymology*; D008969:Molecular Sequence Data; D009154:Mutation; D010088:Oxidoreductases / Q000235:genetics*; D011487:Protein Conformation; D021382:Protein Sorting Signals / Q000032:analysis; D051381:Rats; D015183:Restriction Mapping
chemical_list: D000214:Acyl Coenzyme A; D021382:Protein Sorting Signals; D004247:DNA; D010088:Oxidoreductases