Proteomic analysis of rat liver peroxisome. Presence of peroxisome-specific isozyme of Lon protease

Miki Kikuchi, Naoya Hatano, Sadaki Yokota, Nobuyuki Shimozawa, Tsuneo Imanaka, Hisaaki Taniguchi

Research output: Contribution to journalArticle

209 Citations (Scopus)

Abstract

Subcellular proteomics, which includes isolation of subcellular components prior to a proteomic analysis, is advantageous not only in characterizing large macromolecular complexes such as organelles but also in elucidating mechanisms of protein transport and organelle biosynthesis. Because of the high sensitivity achieved by the present proteomics technology, the purity of samples to be analyzed is important for the interpretation of the results obtained. In the present study, peroxisomes isolated from rat liver by usual cell fractionation were further purified by immunoisolation using a specific antibody raised against a peroxisomal membrane protein, PMP70. The isolated peroxisomes were analyzed by SDS-PAGE combined with liquid chromatography/mass spectrometry. Altogether 34 known peroxisomal proteins were identified in addition to several mitochondrial and microsomal proteins. Some of the latter may reside in the peroxisomes as well. Analysis of membrane fractions identified all known peroxins except for Pex7. Two new peroxisomal proteins of unknown function were of high abundance. One is a bi-functional protein consisting of an aminoglycoside phosphotransferase-domain and an acyl-CoA dehydrogenase domain. The other is a newly identified peroxisome-specific isoform of Lon protease, an ATP-dependent protease with chaperone-like activity. The peroxisomal localization of the protein was confirmed by immunological techniques. The peroxisome-type Lon protease, which is distinct from the mitochondrial isoform, may play an important role in the peroxisomal biogenesis.

Original languageEnglish
Pages (from-to)421-428
Number of pages8
JournalJournal of Biological Chemistry
Volume279
Issue number1
DOIs
Publication statusPublished - Jan 2 2004
Externally publishedYes

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Protease La
Peroxisomes
Liver
Proteomics
Isoenzymes
Rats
Proteins
Organelles
Protein Isoforms
ATP-Dependent Proteases
Kanamycin Kinase
Acyl-CoA Dehydrogenase
Immunologic Techniques
Macromolecular Substances
Cell Fractionation
Mitochondrial Proteins
Protein Transport
Liquid Chromatography
Biosynthesis
Liquid chromatography

ASJC Scopus subject areas

  • Biochemistry

Cite this

Proteomic analysis of rat liver peroxisome. Presence of peroxisome-specific isozyme of Lon protease. / Kikuchi, Miki; Hatano, Naoya; Yokota, Sadaki; Shimozawa, Nobuyuki; Imanaka, Tsuneo; Taniguchi, Hisaaki.

In: Journal of Biological Chemistry, Vol. 279, No. 1, 02.01.2004, p. 421-428.

Research output: Contribution to journalArticle

Kikuchi, Miki ; Hatano, Naoya ; Yokota, Sadaki ; Shimozawa, Nobuyuki ; Imanaka, Tsuneo ; Taniguchi, Hisaaki. / Proteomic analysis of rat liver peroxisome. Presence of peroxisome-specific isozyme of Lon protease. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 1. pp. 421-428.
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