Palmitoylation of neurofascin at a site in the membrane-spanning domain highly conserved among the L1 family of cell adhesion molecules.
This report presents the first evidence that a member of the L1 family of nervous system cell-adhesion molecules is covalently modified by thioesterification with palmitate, and identifies a highly conserved cysteine in the predicted membrane-spanning domain as the site of modification. Neurofascin is constitutively palmitoylated at cysteine-1213 at close to a 1:1 molar stoichiometry. Kinetics of palmitate incorporation into neurofascin expressed in resting neuroblastoma cells indicate that the palmitate modification has the same turnover rate as the polypeptide chain and does not affect the protein stability of neurofascin. Palmitoylation of neurofascin expressed in dorsal root ganglion neurons is not required for delivery of neurofascin to the plasma membrane or targeting to axons. Palmitoylation also has no effect on ankyrin-binding activity of neurofascin, on the oligomeric state of neurofascin in solution, or on cell-adhesion activity of neurofascin expressed in neuroblastoma cells. A significant difference between native and C1213L neurofascin is that these proteins were localized in distinct fractions within a low-density membrane population enriched in signaling molecules. These results indicate a palmitate-dependent targeting of neurofascin to a specialized membrane microdomain.
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Howard Hughes Medical Institute and Department of Cell Biology, Duke University Medical Center, Durham, North Carolina 27710, USA.
Bennett
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D000595:Amino Acid Sequence; D000818:Animals; D017487:Ankyrins / Q000378:metabolism; D015815:Cell Adhesion Molecules / Q000235:genetics* / Q000378:metabolism* / Q000502:physiology; D002460:Cell Line; D017124:Conserved Sequence; D005727:Ganglia, Spinal / Q000166:cytology / Q000378:metabolism; D019811:Hydroxylamine / Q000494:pharmacology; D007700:Kinetics; D008566:Membranes / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation / Q000502:physiology; D009414:Nerve Growth Factors / Q000378:metabolism* / Q000502:physiology; D009474:Neurons / Q000378:metabolism; D019308:Palmitic Acid / Q000037:antagonists & inhibitors / Q000378:metabolism*; D051381:Rats; D013347:Subcellular Fractions / Q000378:metabolism
D017487:Ankyrins; D015815:Cell Adhesion Molecules; D009414:Nerve Growth Factors; C504594:Nfasc protein, rat; D019811:Hydroxylamine; D019308:Palmitic Acid