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 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.
Ramón De Lucas
J
J
Departamento de Microbiología y Parasitología, Facultad de Farmacia, Universidad de Alcalá, Ctra. Madrid-Barcelona Km 33, E-28871 Alcalá de Henares, Madrid, Spain. joser.lucas@uah.es
Martínez
O
O
Pérez
P
P
Isabel López
M
M
Valenciano
S
S
Laborda
F
F
D000906:Antibodies / Q000276:immunology; D001233:Aspergillus nidulans / Q000737:chemistry / Q000166:cytology*; D015153:Blotting, Western; D002352:Carrier Proteins / Q000032:analysis* / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D016253:Microscopy, Immunoelectron; D008928:Mitochondria / Q000737:chemistry* / Q000648:ultrastructure; D020675:Peroxisomes / Q000737:chemistry; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification
D000906:Antibodies; D002352:Carrier Proteins; D011993:Recombinant Fusion Proteins