The Journal of Experimental Medicine · 1988 · 126 citations · 16 references
Life SciencesBiochemistryMolecular BiologyAutoimmunityHepatologyMonoclonal AntibodiesAntibody CharacterizationHepatic PathologyLiver DiseaseHomologyMouse Liver MicrosomesPediatric Autoimmune HepatitisRat Liver MicrosomesRecombinant Cytochrome P450 EnzymesRecombinant Fusion ProteinsRestriction Fragment MappingRestriction Mapping
Children with autoimmune hepatitis have high serum titers of antibodies directed against a 50-kD protein of rat liver endoplasmic reticulum. Affinity-purified anti-50-kD antibodies were used to screen a rat liver cDNA library in lambda GT-11 expression vector. 12 immunopositive clones were obtained. Crossreactivities between fusion proteins of these clones and the 50-kD protein was demonstrated, and four clones were analyzed by restriction mapping, one of them by nucleotide sequencing. Complete identity was found between the restriction maps of two clones (LKMC1 and LKMC2) and that of the 5' end of the rat cytochrome P450 db2. Sequence of a 608-bp fragment of LKMC1 showed complete homology with the rat P450 db2 form. The restriction map of the other two clones (LKMC3 and LKMC4) was identical to that of rat P450 db1. These results suggest that the antigen recognized by LKMA is a P450 of the IID subfamily.
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Haider Raza, Annie John, Abderrahim Nemmar · Cellular Physiology and Biochemistry · 2013 · 47.3K citations · Full text