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. 2015 Feb 11;35(6):2492–2507. doi: 10.1523/JNEUROSCI.4248-14.2015

Table 1.

Identification of phosphorylation sites on mouse brain SV2A

Peptides identified by mass spectrometry Mass Mascot score Possible P sites % Probability (PD1.4 pRS3.1)
>150 kDa
    R.GGLSDGEGPPGGR.G + P (ST) 1234.4310 64 S127 100
    R.GGLSDGEGPPGGRGEAQR.R + P (ST) 1775.6966 35 S127 100
    R.GEGAQDEEEGGASSDATEGHDEDDEIYEGEYQGIPR.A + 2 P (ST) 4000.3955 66 S80, 81 98, 98
    R.GEGAQDEEEGGASSDATEGHDEDDEIYEGEYQGIPR.A + 3 P (ST) 4080.3606 32 S80, S81, T84 100, 100, 100
75–150 kDa
    R.VFSVTHIK.T + P (ST) 1009.4332 44 S393 99
    R.GGLSDGEGPPGGR.G + P (ST) 1234.4322 72 S127 100
    R.GGLSDGEGPPGGRGEAQR.R + P (ST) 1775.6939 50 S127 100
    R.GEGAQDEEEGGASSDATEGHDEDDEIYEGEYQGIPR.A + P (ST) 3920.4297 90 S81 88
    R.GEGAQDEEEGGASSDATEGHDEDDEIYEGEYQGIPR.A + 2 P (ST) 4000.3966 60 S81 and either of S80 or T84 53, 92, 53
    R.GEGAQDEEEGGASSDATEGHDEDDEIYEGEYQGIPR.A + 3 P (ST) 4080.3698 66 S80, S81, T84 100, 100, 100

Endogenous SV2A was immunoprecipitated from mouse brain extract (8 mg). The immunoprecipitates were electrophoresed on a polyacrylamide gel, and after Colloidal blue staining, bands corresponding to SV2A were excised and digested with trypsin. Phosphorylated peptides were identified by LC-MS/MS analysis. The data analysis was as in Materials and Methods. The sequence of each peptide is shown with possible phosphorylation sites underlined. Also shown are the mass of the peptide, the Mascot score [of the peptide identification, in which individual ion scores >21 indicate identity or extensive homology (p < 0.05)], and the percentage probability of the phosphosite assignment, as determined by the phosphoRS 3.1 component of the Proteome Discoverer 1.4 software.