Table 4.
DAB Neuroproteome Relative abundance of brain proteins of young adult C57/BL6 mice 168 hours after a single intraperitoneal injection of 1,2-DAB (50 mg/kg body weight), 1,3-DAB (50 mg/kg), or DAB Vehicle (2% acetone in saline). Half brains were pooled from two animals from each group. Each sample was homogenized in 1 ml 100 mM ammonium bicarbonate, 5 mM dithiothreitol, pH 7.8, centrifuged at 80K for 1h in a TL-100 rotor, the supernatant collected and assayed for protein at a 1:100 dilution. 100 mcg of each sample was digested with 1:50 trypsin to protein (w/w), for 1h at 37oC (20% ACN). Peptide clean-up was performed on a SP-C18 column and eluted in 80% acetonitrile (CAN), 0.1% acetic acid, and 0.01% trifluoroacetic acid. Each sample was run in duplicate on a 9-tesla Fourier-transform ion cyclotron resonance-mass spectrometer. The mouse brain voxel PMT database was used for peak matching of proteins of differing abundance. A total of 1643 redundant proteins was identified, of which 548 proteins were modulated by 1,2-DAB, with 196 mapped to cell networks. Performed in collaboration with J. Pounds and J. Adkins, Pacific Northwest National Laboratory, U.S. Department of Energy Richland, Washington.
| PROTEIN ABUNDANCE | Change in Abundance | Relative Abundance | |||
|---|---|---|---|---|---|
| PROTEIN NAME/FUNCTION | 1,2-DAB/1,3-DAB | 1,2-DAB/Vehicle | 1,2-DAB | 1,3-DAB | Vehicle |
| MICROTUBULE-INTERACTING | |||||
| Stathmin | 6.98 | 8.85 | 108.54 | 15.55 | 12.27 |
| oral-facial-digital syndrome 1 | 4.68 | ∞ | 46.46 | 9.96 | 0.00 |
| restin (clip-170, clip-50) | 3.38 | 5.71 | 8.62 | 2.55 | 1.51 |
| microtubule-associated protein 2 | 5.47 | 3.79 | 3.94 | 0.72 | 1.04 |
| Synuclein (alpha + beta) | 4.12 | 2.09 | 2.72 | 0.66 | 1.30 |
| OTHER STRUCTURAL PROTEINS | |||||
| neurofilament medium (NF-M) | 4.09 | 7.37 | 2.21 | 0.54 | 0.30 |
| neurofilament light (NF-L) | 1.64 | 1.24 | 0.46 | 0.28 | 0.37 |
| actin alpha 1 | 2.30 | 1.60 | 18.77 | 8.15 | 11.73 |
| actin alpha 2 | 14.85 | 50.77 | 15.74 | 1.06 | 0.31 |
| procollagen type V1 alpha 2 | 3.29 | 5.38 | 5.16 | 1.57 | 0.96 |
| Peripherin | 3.84 | 2.32 | 1.23 | 0.32 | 0.53 |
| CELLULAR ENERGY METABOLISM | |||||
| malate dehydrogenase, soluble | 23.59 | 75.19 | 24.06 | 1.02 | 0.32 |
| triosephosphate isomerase | 2.20 | 3.97 | 18.10 | 8.22 | 4.56 |
| enolase-2 (neuronal) | 1.73 | ∞ | 14.92 | 8.64 | 0.00 |
| NADH dehydrogenase Fe-S protein 3 | 0.31 | 0.45 | 10.93 | 35.83 | 24.13 |
| phosphoglycerate kinase 2 | 5.34 | 10.69 | 7.48 | 1.40 | 0.70 |
| ATP synthase beta-chain | 3.17 | 5.07 | 0.76 | 0.24 | 0.15 |
| VESICULAR TRACKING | |||||
| synapsin 1 | ∞ | 2.65 | 11.46 | 0.00 | 4.33 |
| kinesin family member 20A | 38.13 | ∞ | 6.10 | 0.16 | 0.00 |