Mechanisms and Consequences of Dopamine Depletion-Induced Attenuation of the Spinophilin/Neurofilament Medium Interaction

Figure 4

Synaptic spinophilin phosphorylation at Ser17 and Ser100 is enhanced by DA depletion. The substantia nigra of adult C57Bl6 mice was unilaterally lesioned with 6-OHDA. Striata from the intact and lesioned hemispheres were collected, fractionated into a membrane and synaptic fraction, and immunoprecipitated for spinophilin as in Figure 1. The region of the gel containing spinophilin and NF-M (Figure 1(a)) was excised and analyzed by mass spectrometry. (a) MS/MS spectra of a spinophilin tryptic fragment containing phosphorylated Ser17. (b) Quantification of the normalized AUC of the XIC matching the Ser17 in lesion and intact tissue normalized to the corresponding intact value. (c) MS/MS spectra of a spinophilin tryptic fragment containing phosphorylated Ser100. (d) Quantification of the normalized AUC of the XIC matching the Ser100 in lesion and intact tissue normalized to the corresponding intact value. compared to the intact group. . Mass spectrometry was performed on an Orbitrap Velos mass spectrometer.

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