A novel cell model to study the function of the adrenoleukodystrophy-related protein.

Abstract

X-linked adrenoleukodystrophy (X-ALD) is a neurodegenerative disorder due to mutations in the ABCD1 (ALD) gene. ALDRP, the closest homolog of ALDP, has been shown to have partial functional redundancy with ALDP and, when overexpressed, can compensate for the loss-of-function of ALDP. In order to characterize the function of ALDRP and to understand the phenomenon of gene redundancy, we have developed a novel system that allows the controlled expression of the ALDRP-EGFP fusion protein (normal or non-functional mutated ALDRP) using the Tet-On system in H4IIEC3 rat hepatoma cells. The generated stable cell lines express negligible levels of endogenous ALDRP and doxycycline dosage-dependent levels of normal or mutated ALDRP. Importantly, the ALDRP-EGFP protein is targeted correctly to peroxisome and is functional. The obtained cell lines will be an indispensable tool in our further studies aimed at the resolution of the function of ALDRP to characterize its potential substrates in a natural context.

• Authors: Fabien Gueugnon, Natalia Volodina, Jaoued Et Taouil, Tatiana E Lopez, Catherine Gondcaille, Anabelle Sequeira-Le Grand, Petra A W Mooijer, Stephan Kemp, Ronald J A Wanders, Stéphane Savary
• Journal: Biochemical and biophysical research communications (Biochem Biophys Res Commun) Vol. 341 Issue 1 Pg. 150-7 (Mar 03 2006) ISSN: 0006-291X [Print] United States
• PMID: 16412981 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• ABCD2 protein, human
• ATP Binding Cassette Transporter, Subfamily D
• ATP-Binding Cassette Transporters
• Recombinant Fusion Proteins

Topics

• ATP Binding Cassette Transporter, Subfamily D
• ATP-Binding Cassette Transporters (genetics, metabolism)
• Adrenoleukodystrophy (genetics, metabolism)
• Animals
• Carcinoma, Hepatocellular (genetics, metabolism)
• Cell Line, Tumor
• Disease Models, Animal
• Protein Engineering (methods)
• Rats
• Recombinant Fusion Proteins (metabolism)
• Transfection (methods)