In addition, the chaperone protein, Stress MYO10
Protein-70, which is involved in the translocation and folding of nascent polypeptide chains of mitochondrial origin, was also down-regulated (Table?1; Figs?1 and 2). Neural cell adhesion molecule 120kDa, a glycosylphosphatidylinositol-linked adhesion protein, predominantly expressed in OLs (Goridis and Brunet 1992), was increased in CGTnull-myelin, with no alteration in CSTnull-myelin (Table?1; Figs?1 and 2a). While several proteins were altered exclusively in CGTnull-myelin, the only protein altered exclusively in CSTnull-myelin was SirT2. The increase was only in its minor variant-1 but not in the more abundant variant-2 (Table?1; Figs?1 and 2e). SirT2, involved in the regulation of cellular differentiation, is expressed in OLs and myelin (Taylor et?al. 2004; Roth et?al. 2005; Werner et?al. 2007), and is down-regulated in PLPnull myelin (Werner et?al. 2007). Among the similarly altered proteins in CGTnull and CSTnull-myelin, the most striking was the reduction of Septin-2, -4, and -8 observed XL765 nmr
by both 2D-DIGE and immunoblotting (Table 1; Figs?1 and 2g-i). All three Septins were enriched in MB subfraction corresponding to a compact myelin compartment (Menon et?al. 2003). Remarkably, Septin-8 (variant-2) was almost exclusively found in the compact myelin fraction of WT mice (Fig.?2g). Further, it was decreased fourfold (p?<?0.02; t-test) in CGTnull and twofold (p?<?0.05; t-test) in CSTnull-myelin. Likewise, Septin-2 and -4 were also reduced twofold in the same myelin subfraction observed by either 2D-DIGE (Table 1; Fig.?1) or immunoblotting (Fig.?2h and i). Although 2D-DIGE had identified alterations in Septin-2, -4, and -8 in <a href="http://www.selleckchem.com/products/yap-tead-inhibitor-1-peptide-17.html
">selleck inhibitor the mutant myelin, we further investigated for possible changes in other Septins by immunoblotting of myelin subfractions of mutant mice and WT littermates (Supporting information Fig.?S2a�Ch). Unlike Septin-2, -4, and -8, the other Septins examined (Septin-5, -6, -7, -11) showed no enrichment in the MB subfraction of WT myelin over total brain homogenates, suggesting that these Septins were not enriched in the compact myelin compartment. Nevertheless, the already low level of these Septins in this compartment was further reduced in the two mutants. Septin-5 was unaffected. Together, these results suggest that the Septin family members are distributed differently in normal myelin compartments and that their expression is regulated by galactolipids. Ezrin was another protein that was reduced in both mutants observed by 2D-DIGE and confirmed by immunoblot analysis (Table?1; Figs?1 and 2f). Interestingly, the down-regulation was exclusively in the MB and dispersion but not in the heavy-band and pellet subfractions suggesting that this regulation in restricted to the compact myelin compartment.