The Aspergillus nidulans carnitine carrier encoded by the acuH gene is exclusively located in the mitochondria

The location of the Aspergillus nidulans carnitine/acyl-carnitine carrier (ACUH) was studied. ACUH with a His-tag at its N-terminus was over-expressed in Escherichia coli and purified by Ni 2+ affinity chromatography. The purified protein was utilised to raise polyclonal antibodies which were charac...

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Bibliographic Details
Published in:FEMS microbiology letters 2001-07, Vol.201 (2), p.193-198
Main Authors: Ramón De Lucas, J., Martı́nez, Oscar, Pérez, Patricia, Isabel López, M., Valenciano, Susana, Laborda, Fernando
Format: Article
Language:English
Subjects:
acuH gene
affinity chromatography
alcA gene
Antibodies - immunology
Aspergillus nidulans
Aspergillus nidulans - chemistry
Aspergillus nidulans - cytology
Aspergillus nidulans carnitine/acyl-carnitine carrier
Bacteriology
Biological and medical sciences
Blotting, Western
carnitine
Carnitine carrier
carnitine/acyl-carnitine carrier
Carrier Proteins - analysis
Carrier Proteins - genetics
Carrier Proteins - immunology
Carrier Proteins - isolation & purification
Cloning, Molecular
Escherichia coli
Escherichia coli - genetics
Fatty acid
Fundamental and applied biological sciences. Psychology
genes
Genetics
Growth, nutrition, metabolism, transports, enzymes. Molecular biology
Microbiology
Microscopy, Immunoelectron
mitochondria
Mitochondria - chemistry
Mitochondria - ultrastructure
Mitochondrion
Mycology
nickel
Peroxisome
Peroxisomes - chemistry
polyclonal antibodies
Protein Transport
Recombinant Fusion Proteins - chemistry
Recombinant Fusion Proteins - immunology
Recombinant Fusion Proteins - isolation & purification
Transport
Western blotting
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Summary:The location of the Aspergillus nidulans carnitine/acyl-carnitine carrier (ACUH) was studied. ACUH with a His-tag at its N-terminus was over-expressed in Escherichia coli and purified by Ni 2+ affinity chromatography. The purified protein was utilised to raise polyclonal antibodies which were characterised by Western blotting. For localisation studies A. nidulans T1 strain, that contains the acuH gene under control of the strong promoter alcA p, was derived. Results obtained demonstrate the exclusively mitochondrial localisation of ACUH and therefore exclude the targeting of the acuH gene product to the peroxisomal membrane.
ISSN:0378-1097
1574-6968
DOI:10.1016/S0378-1097(01)00261-0