Translocation of Acyl-CoA Oxidase into Peroxisomes Requires ATP Hydrolysis but Not a Membrane Potential

• Published in: The Journal of cell biology 1987-12, Vol.105 (6), p.2915-2922
• Main Authors: Imanaka, Tsuneo, Small, Gillian M., Lazarow, Paul B.
• Format: Article
• Language: English
• Subjects: Acyl-CoA Oxidase; Adenosine Triphosphate - metabolism; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Biological Transport, Active; Carbonyl Cyanide m-Chlorophenyl Hydrazone - pharmacology; Cell Fractionation; Centrifugation, Density Gradient; Chloroplasts; Enzymes; Enzymes and enzyme inhibitors; Fundamental and applied biological sciences. Psychology; Imports; In Vitro Techniques; Iohexol; Ionophores - pharmacology; Kinetics; Liver; Magnesium - pharmacology; Magnesium Chloride; Membrane Potentials; Messenger RNA; Microbodies - metabolism; Mitochondria; Organelles; Oxidases; Oxidoreductases; Oxidoreductases - metabolism; Peroxisomes; Potassium Chloride - pharmacology; Protein transport; Temperature
• ISSN: 0021-9525; 1540-8140
• DOI: 10.1083/jcb.105.6.2915

An efficient system for the import of newly synthesized proteins into highly purified rat liver peroxisomes was reconstituted in vitro. 35 S-Labeled acyl-CoA oxidase (AOx) was incorporated into peroxisomes in a proteinase K-resistant fashion. This import was specific (did not occur with mitochondria) and was dependent on temperature, time, and peroxisome concentration. Under optimal conditions ∼30% of [35 S] AOx became proteinase resistant. The import of AOx into peroxisomes could be dissociated into two steps: (a) binding occurred at 0°C in the absence of ATP; (b) translocation occurred only at 26°C and required the hydrolysis of ATP. GTP would not substitute for ATP and translocation was not inhibited by carbonylcyanide-m-chlorophenylhydrazone, valinomycin, or other ionophores.