Chemical deacylation reduces the adhesive properties of proteolipid protein and leads to decompaction of the myelin sheath

Myelin proteolipid protein (PLP) contains thioester-bound, long-chain fatty acids which are known to influence the structure of the molecule. To gain further insights into the role of this post-translational modification, we studied the effect that chemical deacylation of PLP had on the morphology o f myelin and on the protein's ability to mediate the clustering of lipid ve sicles. Incubation of rat optic nerves in isoosmotic solutions containing 1 00 mm hydroxylamine (HA) pH 7.4 led to deacylation of PLP and decompaction of myelin lamellae at the level of the intraperiod line. Incubation of nerv es with milder nucleophilic agents (Tris and methylamine) or diluted HA, co nditions that do not remove protein-bound fatty acids, caused no alteration s in myelin structure. Other possible effects of HA which could have affect ed myelin compaction indirectly were ruled out. Incubation of optic nerves with 50 mm dithioerythritol (DTE) also led to the splitting of the myelin i ntraperiod line and this change again coincided with the removal of fatty a cids. In addition, the apparently compacted CNS myelin in the PLP-less myel in-deficient rat, like that in tissue containing deacylated PLP, was readil y decompacted upon incubation in isoosmotic buffers, suggesting that the fu nction of PLP as a stabilizer of the interlamellar attachment is, at least in part, mediated by fatty acylation. Furthermore, in contrast to the nativ e protein, PLP deacylated with either HA or DTE failed to induce the cluste ring of phosphatidylcholine/cholesterol vesicles in vitro. This phenomenon is not due to side-effects of the deacylation procedure since, upon partial repalmitoylation, the protein recovered most of its original vesicle-clust ering activity. Collectively, these findings suggest that palmitoylation, b y influencing the adhesive properties of PLP, is important for stabilizing the multilamellar structure of myelin.