PTS2 protein import into mammalian peroxisomes.

Abstract

Peroxisome targeting signal (PTS)2 directs proteins from their site of synthesis in the cytosol to the lumen of the peroxisome. Unlike PTS1 which is present in the great majority of peroxisomal matrix proteins and whose import mechanics have been dissected in considerable detail, PTS2 is a relatively rare topogenic signal whose import mechanisms are far less well understood. However, as is the case for PTS1 proteins, an inability to import PTS2 proteins leads to human disease. In this report, we describe the biochemical characterization of mammalian PTS2 protein import using a semi-permeabilized cell system. We show that a PTS2-containing reporter molecule is taken up by peroxisomes in a reaction that is time-, temperature-, ATP-, and cytosol-dependent. Furthermore, the import process is specific, saturable, and requires action of the chaperone Hsc70, the cochaperone Hsp40, and the peroxins Pex5p and Pex14p. We also demonstrate peroxisomal translocation of PTS2 reporter/antibody complexes confirming the import competence of higher order structures. Importantly, cultured fibroblasts from patients with the rhizomelic form of chondrodysplasia punctata (RCDP) which are deficient for the PTS2 receptor protein, Pex7p, are unable to import the PTS2 reporter in this assay. The ability to monitor PTS2 import in vitro will permit, for the first time, a detailed comparison of the biochemical properties of PTS1 and PTS2 protein import.

• Authors: J E Legakis, S R Terlecky
• Journal: Traffic (Copenhagen, Denmark) (Traffic) Vol. 2 Issue 4 Pg. 252-60 (Apr 2001) ISSN: 1398-9219 [Print] England
• PMID: 11285135 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• DNAJB1 protein, human
• HSC70 Heat-Shock Proteins
• HSP40 Heat-Shock Proteins
• HSP70 Heat-Shock Proteins
• HSPA8 protein, human
• Heat-Shock Proteins
• Molecular Chaperones
• Peroxisomal Targeting Signal 2 Receptor
• Receptors, Cytoplasmic and Nuclear
• Recombinant Fusion Proteins
• Adenosine Triphosphate
• Acetyl-CoA C-Acetyltransferase

Topics

• Acetyl-CoA C-Acetyltransferase (genetics, metabolism)
• Adenosine Triphosphate (metabolism)
• Animals
• CHO Cells
• Cells, Cultured
• Chondrodysplasia Punctata, Rhizomelic (genetics, metabolism, pathology)
• Cricetinae
• Cytosol (metabolism)
• Fibroblasts
• Fluorescent Antibody Technique
• HSC70 Heat-Shock Proteins
• HSP40 Heat-Shock Proteins
• HSP70 Heat-Shock Proteins (metabolism)
• Heat-Shock Proteins (metabolism)
• Humans
• Molecular Chaperones (metabolism)
• Peroxisomal Targeting Signal 2 Receptor
• Peroxisomes (metabolism)
• Protein Transport
• Receptors, Cytoplasmic and Nuclear (deficiency, genetics, metabolism)
• Recombinant Fusion Proteins (chemistry, genetics, metabolism)
• Substrate Specificity
• Temperature
• Time Factors