Structural and immunological characterization of the amino-terminal domain of mammalian neural cell adhesion molecules.
The neural cell adhesion molecules (N-CAMs) are a group of structurally and immunologically related glycoproteins found in vertebrate neural tissues. Adult brain N-CAMs have apparent molecular weights of 180,000 and 140,000 with an additional form at 120,000 in murine brain. In embryonic brain, N-CAMs are represented by a highly sialylated form with an apparent molecular weight greater than 180,000. We have used monoclonal antibodies that cross-react with N-CAMs of various mammalian species to purify N-CAMs from adult murine and bovine brains and from embryonic murine brains. We determined the amino acid sequences of the amino-terminal domains of these molecules: Leu-Gln-Val-Asp-Ile-Val-Pro-Ser-Gln-Gly-Glu-Ile-Ser-Val-Gly-Glu-Ser. This sequence is highly conserved among all three forms of adult murine brain N-CAM as well as embryonic murine brain N-CAM and adult bovine brain N-CAM. Based on this sequence, we synthesized an undecapeptide and used it to raise a site-directed polyclonal antiserum. This antiserum reacted with the intact N-CAM in liquid phase radioimmunoassays, immunoblotting experiments, and immunofluorescent labeling of cells. The antiserum also reacted with N-CAMs in extracts of brain tissues from different species, confirming the highly conserved nature of the amino-terminal domain of mammalian N-CAMs. Immunofluorescence experiments indicated that this domain resides on the outer surfaces of cells that express N-CAMs, in both primary neuronal cell culture and in cell lines.
D000595:Amino Acid Sequence; D000818:Animals; D000954:Antigens, Surface / Q000032:analysis* / Q000276:immunology; D001923:Brain Chemistry; D002417:Cattle; D015815:Cell Adhesion Molecules; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D007163:Immunosorbent Techniques; D051379:Mice; D008856:Microscopy, Fluorescence; D008970:Molecular Weight
D000954:Antigens, Surface; D015815:Cell Adhesion Molecules