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. 2011 Jul;134(Pt 7):e178.
doi: 10.1093/brain/awr015. Epub 2011 Feb 24.

Salivary α-synuclein and DJ-1: potential biomarkers for Parkinson's disease

Ivana Devic, Hyejin Hwang, John Scott Edgar, Kenneth Izutsu, Richard Presland, Catherine Pan, David R Goodlett, Yu Wang, Jeff Armaly, Vitor Tumas, Cyrus P Zabetian, James B Leverenz, Min Shi, Jing Zhang

Salivary α-synuclein and DJ-1: potential biomarkers for Parkinson's disease

Ivana Devic et al. Brain. 2011 Jul.
No abstract available

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Figures

Figure 1
Figure 1
Characterization of α-Syn and DJ-1 in human whole saliva. (A) To determine the presence of α-Syn in saliva, a pooled saliva sample was analysed by immunoblotting with a rabbit anti-human α-Syn antibody ASY-1. The specificity of the detected band was confirmed by an immunodepletion experiment with a BD (Becton, Dickinson and Company) Biosciences mouse anti-α-Syn antibody. Lane 1 from the left: 1 ng human α-Syn standard (STD); Lane 2, 100 µg of protein from saliva supernatant (SP); Lane 3, 100 µg of proteins from saliva after immunodepletion; Lane 4, 100 µg of protein from immunoprecipitated/enriched saliva by the BD antibody; Lane 5, 100 µg of protein from enriched saliva by normal mouse IgG. A distinct α-Syn band at 15 kDa was detected in saliva. A lower and likely non-specific band was also detected. (B) The 15 kDa band detected by western blotting was further confirmed to be α-Syn by in-gel digestion and mass spectrometry. Shown is the fragmentation spectrum of mass spectrometry/mass spectrometry for ion 2740.43 of α-Syn. The sequence shown is TVEGAGSIAAATGFVK, with y- and b-series ions interpreted. The results demonstrate that three fragments, corresponding to a total of 47 residues of the 140 amino acids, of full-length α-Syn were identified (see also Supplementary Fig. 2 and Table 2). (C) Similarly, the pooled saliva sample was also analysed by immunoblotting with a polyclonal goat antibody against DJ-1. The specificity of the detected band was confirmed by an immunodepletion experiment with a polyclonal rabbit antibody. (D) The 25 kDa band detected by western blotting was further confirmed to be DJ-1 by in-gel digestion and mass spectrometry. Shown is the fragmentation spectrum of mass spectrometry/mass spectrometry for ion 2766.30 of DJ-1. The sequence shown is DVVICPDASLEDAK, with y- and b-series ions interpreted. The results demonstrate that nine fragments, corresponding to a total of 86 residues of the 189 amino acids, of full-length DJ-1 were identified (see also Supplementary Fig. 2 and Table 2).
Figure 2
Figure 2
Salivary α-Syn and DJ-1 levels in patients with Parkinson's disease and healthy controls. α-Syn and DJ-1 protein levels in saliva were measured by Luminex in healthy controls (CTRL) and patients with Parkinson's disease (PD). The α-Syn levels (A) tended to decrease while the DJ-1 levels (C) tended to increase in the Parkinson's disease compared with the control group, whether normalized to the saliva total protein level or not. In a subset of the Parkinson's disease subjects with known UPDRS motor scores, the correlation between the UPDRS scores and salivary α-Syn (B) or DJ-1 (D) levels was also examined. α-Syn levels appeared to be negatively correlated with severity of motor symptoms, whether normalized (red rectangle) or not (black circle), while DJ-1 levels did not correlate well with UPDRS motor scores. UPDRS = Unified Parkinson's Disease Rating Scale.
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