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J Biol Chem, Vol. 274, Issue 12, 8224-8230, March 19, 1999
From the In the central nervous system, many cell adhesion
molecules are known to participate in the establishment and remodeling
of the neural circuit. Some of the cell adhesion molecules are known to
be anchored to the membrane by the glycosylphosphatidylinositol (GPI)
inserted to their C termini, and many GPI-anchored proteins are known
to be localized in a Triton-insoluble membrane fraction of low density
or so-called "raft." In this study, we surveyed the GPI-anchored
proteins in the Triton-insoluble low density fraction from 2-week-old
rat brain by solubilization with phosphatidylinositol-specific phospholipase C. By Western blotting and partial peptide sequencing after the deglycosylation with peptide N-glycosidase F, the
presence of Thy-1, F3/contactin, and T-cadherin was shown. In addition, one of the major proteins, having an apparent molecular mass of 36 kDa
after the peptide N-glycosidase F digestion, was found to
be a novel protein. The result of cDNA cloning showed that the
protein is an immunoglobulin superfamily member with three C2 domains
and has six putative glycosylation sites. Since this protein shows high
sequence similarity to IgLON family members including LAMP, OBCAM,
neurotrimin, CEPU-1, AvGP50, and GP55, we termed this protein Kilon (a
kindred of IgLON). Kilon-specific monoclonal
antibodies were produced, and Western blotting analysis showed that
expression of Kilon is restricted to brain, and Kilon has an apparent
molecular mass of 46 kDa in SDS-polyacrylamide gel electrophoresis in
its expressed form. In brain, the expression of Kilon is already
detected in E16 stage, and its level gradually increases during
development. Kilon immunostaining was observed in the cerebral cortex
and hippocampus, in which the strongly stained puncta were observed on
dendrites and soma of pyramidal neurons.
Characterization of a Novel Rat Brain
Glycosylphosphatidylinositol-anchored Protein (Kilon), a Member of the
IgLON Cell Adhesion Molecule Family
§,,§,,,,, and
Department of Biotechnology,
§ Venture Laboratory, Kyoto Institute of Technology,
Kyoto 606-8585, Japan
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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