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. Author manuscript; available in PMC: 2012 Jul 14.
Published in final edited form as: Neuron. 2011 Jul 14;71(1):131–141. doi: 10.1016/j.neuron.2011.05.033

Figure 6. Palmitoylation regulates PSD-95 nitrosylation.

Figure 6

(A) 2-BP treatment of HEK-nNOS cells increases nitrosylation. HEK-nNOS cells expressing PSD-95-1-433-FLAG or the corresponding C3, C5S mutant were treated with 100 μM 2-BP for 4 hr and analyzed by biotin switch. β-tubulin is a control nitrosylated protein unaffected by 2BP treatment. Left, representative result. Right, quantification of nitrosylated PSD-95-1-433-FLAG, analyzed as in Figure 2. n=3, *, p < 0.02 relative to untreated.

(B) ZDHHC8−/− mice have increased endogenous SNO-PSD-95. Brain homogenates from age-matched wt or ZDHHC8−/− mice were analyzed by biotin switch and ABE. Left, representative result. Middle, quantification for the biotin switch, analyzed as in (A). n=3, *, p < 0.04 relative to wild type. Right, quantification for ABE, n=3, *, p < 0.05 relative to wild type. GAPDH nitrosylation is also demonstrated but does not increase in the ZDHHC−/− mouse.

(C) Basal nitrosylation of NR2A was demonstrated in wild type and ZDHHC8−/− mouse brain by the biotin switch assay. Palmitoylation was measured by the ABE assay.

(D) Decreased binding of NR2B to PSD-95 in ZDDHC−/− mice. NR2B was immunoprecipitated from ZDHHC−/− or wild type brain homogenate and blotted for PSD-95.