Limkain b1, a novel human autoantigen localized to a subset of ABCD3 and PXF marked peroxisomes

Abstract

Detection of self-reactive antibodies has an established role in the diagnosis and monitoring of many human autoimmune diseases. Autoantibodies with restricted reactivity to cytoplasmic compartments and structures are an occasional incidental finding following routine examination of serum for antinuclear antibody reactivity. A prerequisite for rational exploitation of self-reactive antibodies, in either clinical or research settings, is the establishment of the molecular identity of the target autoantigen(s). Here we report on the identification of a novel autoantigen that co-localizes with a subset of cytoplasmic microbodies marked by ABCD3 (PMP-70) and/or PXF (PEX19). Immunoscreening a HeLa cell cDNA expression library with a human autoimmune serum identified two clones that encode fragments of limkain b1 (LKAP). We demonstrate that mouse polyclonal antibodies raised against a bacterially expressed fragment of limkain b1 mark the same cytoplasmic structures as human serum, as does an EGFP:LKAPCT429 fusion protein expressed in HeLa cells. An immunoblot screen against a bacterially expressed MBP:LKAPCT429 fusion protein substrate, using a cohort of 16 additional human sera that display Hep 2 cell cytoplasmic staining patterns similar to the prototype serum, identified three additional sera reactive to limkain b1. This is the first report establishing the molecular identity of a peroxisomal autoantigen. Preliminary results suggest that limkain b1 may be a relatively common target of human autoantibodies reactive to cytoplasmic vesicle-like structures.