Introduction
Methods
General overview of the methodology
Sample procurement, protein extraction, and quantitation from CSF of deceased AD patients
Patient selection, sample procurement, and protein preparation from blood serum of AD patients
Sample procurement and protein preparation from blood samples of patients with unknown AD diagnosis
Cy-dye labeling and 2D gel electrophoresis for differential protein expression analysis
Analysis of differential protein expression from CSF samples
Protein identification by mass spectrometry
Two-dimensional multiplexed Western blotting
LC-MS/MS-based quantitative analysis of ApoE
Results
Proteomic analysis of CSF shows that pI distributions are altered in AD patients as a function of the APOE genotype
Spot number | Protein name/Protein ID/Uniprot Accession Number/RefSeq | 33 N versus 33D | 33 N versus 44D | N versus D | Molecular weight | pI |
---|---|---|---|---|---|---|
429 | Serum albumin, Chain A; ALBU_HUMAN P02768/NP_000468.1 | -1.29 | -1.37 | -1.33 | 69,366.68 | 5.92 |
P = 0.00034 | ||||||
541 | Transferrin variant; Q53H26_HUMAN Q53H26 /IP|00022463 | -1.53 | -1.50 | -1.51 | 77,079.85 | 6.68 |
P = 0.0011 | ||||||
575 | alpha 1-B glycoprotein; A1BG_HUMAN P04217/NP_570602.2 | -1.22 | -1.18 | -1.20 | 54,272.56 | 5.58 |
P = 0.0013 | ||||||
579 | alpha 1-B glycoprotein; A1BG_HUMAN P04217/NP_570602.2 | -1.31 | -1.31 | -1.31 | 54,272.56 | 5.56 |
P = 0.0015 | ||||||
614 | Transferrin variant; Q53H26_HUMAN Q53H26/IP|00022463 | +1.70 | +1.64 | +1.67 | 770,79.85 | 7.04 |
P = 0.0024 | ||||||
990 | Apolipoprotein H (beta-2-glycoprotein I) APOH_HUMAN P02749/NP_000033.2 | +1.67 | +1.60 | +1.63 | 38,298.16 | 8.34 |
P = 0.0021 | ||||||
1000 | Tubulin beta-2A chain; TBB2A_HUMAN Q13885/NP_001060.1 | +1.29 | +1.73 | +1.51 | 49,906.67 | 4.78 |
P = 0.0058 | ||||||
1008 | Keratin, type II cytoskeletal 1; K2C1_HUMAN P04264/NP_006112.3 | +2.94 | +2.62 | +2.77 | 66,038.73 | 8.15 |
P = 0.002 | ||||||
1017 | alpha-1-antitrypsin (SerpinA1); A1AT_HUMAN P01009/NP_000286.3 | +1.86 | +2.26 | +2.06 P = 0.0021 | 46,739.55 | 5.37 |
1118 | Pigment epithelium-derived factor (SerpinF1); PEDF_HUMAN P36955/NP_002606.3 | +1.54 | +1.99 | +1.77 | 46,342.30 | 5.97 |
P = 7.4E-05 | ||||||
1188 | ALB protein (growth-inhibiting protein 20), Isoform 2; ALBU_HUMAN P02768/NP_000468.1 | +1.26 | +1.23 | +1.24 | 47,360.49 | 5.97 |
P = 0.0065 | ||||||
1268 | Glutamine synthetase; GLNA_HUMAN; P15104/NP_001028216.1 | -1.38 | -1.55 | -1.46 | 42,064.46 | 6.43 |
P = 0.0025 | ||||||
1278 | Creatine kinase B-type; KCRB_HUMAN; P12277/NP_001814.2 | -1.46 | -1.40 | -1.43 | 42,644.28 | 5.34 |
P = 0.0077 | ||||||
1324 | Fructose-bisphosphate aldolase A; ALDOA_HUMAN P04075/NP_000025.1 | -1.49 | -1.53 | -1.51 | 39,420.02 | 8.30 |
P = 0.0074 | ||||||
1332 | Aspartate aminotransferase, cytoplasmic; AATC_HUMAN; P17174/NP_002070.1 | -1.28 | -1.49 | -1.38 | 46,247.51 | 6.53 |
P = 0.0099 | ||||||
1363 | Aspartate aminotransferase, cytoplasmic; AATC_HUMAN; P17174/NP_002070.1 | -1.27 | -1.41 | -1.33 | 46,247.51 | 6.50 |
P = 0.0003 | ||||||
1451 | Glyceraldehyde-3-phosphate dehydrogenase G3P_HUMAN P04406/NP_002037.2 | -2.05 | -1.75 | -1.87 | 36,042.22 | 8.57 |
P = 0.0002 | ||||||
1454 | Glyceraldehyde-3-phosphate dehydrogenase G3P_HUMAN P04406/NP_002037.2 | -1.45 | -1.42 | -1.44 | 36,042.22 | 8.59 |
P = 0.013 | ||||||
1469 | Glyceraldehyde-3-phosphate dehydrogenase G3P_HUMAN P04406/NP_002037.2 | -1.58 | -1.52 | -1.55 | 36,042.22 | 8.63 |
P = 0.00094 | ||||||
1470 | Glyceraldehyde-3-phosphate dehydrogenase G3P_HUMAN P04406/NP_002037.2 | -1.80 | -1.72 | -1.75 | 36,042.22 | 8.67 |
P = 0.0005 | ||||||
1472 | Clusterin; Apolipoprotein J; Complement-associated protein SP-40; CLUS_HUMAN; P10909/NP_001164609.1 | +2.01 | +2.01 | +2.01 | 50,062.56 | 5.89 |
P = 5.10E-05 | ||||||
1521 | Apolipoprotein E4; APOE_HUMAN P02649/NP_000032.1 | +2.14 | +3.34 | +2.74 | 36,154.08 | 5.65 |
P = 0.00023 | ||||||
1523 | Clusterin; Apolipoprotein J; Complement associated protein SP-40; CLUS_HUMAN; P10909/NP_001164609.1 | +1.78 | +2.17 | +1.97 | 50,062.56 | 5.89 |
P = 3.50E-06 | ||||||
1527 | Apolipoprotein E4; APOE_HUMAN P02649/NP_000032.1 | +2.02 | +2.42 | +2.22 | 36,154.08 | 5.65 |
P = 1.30E-05 | ||||||
1535 | Complement component 4A; C4A; Q5JNX2_HUMAN; Q5JNX2/IP|00643525 | +1.85 | +1.94 | +1.89 | ~3,5000 | 6.43 |
P = 7.10E-05 | ||||||
1554 | Transthyretin; TTHY_HUMAN; P02766/NP_000362.1 | +2.05 | +2.58 | +2.32 | 15,887.03 | 5.52 |
P = 0.00078 | ||||||
1779 | Ig Kappa chain C region; IGKC; IGKC_HUMAN; P01834/IP|00909649 | -1.98 | -2.14 | -2.06 | 11,608.86 | 5.58 |
P = 0.0061 | ||||||
1845 | Prostaglandin-H2 D-isomerase; cerebrin-28; PTGDS_HUMAN; P41222/NP_000945.3 | +1.45 | +1.51 | +1.48 | 21,028.82 | 7.66 |
P = 0.0077 | ||||||
1912 | Apolipoprotein A-1; ApoA1; APOA1-HUMAN; P02647/NP_000030.1 | -1.78 | -1.84 | -1.81 | 30,777.83 | 5.56 |
P = 0.0037 | ||||||
1982 | Superoxide dismutase [Mn], mitochondrial; SODM_HUMAN; P04179/NP_000627.2 | +1.66 | +1.62 | +1.64 | 24,722.09 | 8.35 |
P = 0.0011 | ||||||
2067 | Apolipoprotein A-1; ApoA1; APOA1-HUMAN; P02647/NP_000030.1 | -1.95 | -2.09 | -2.02 | 30,777.83 | 5.56 |
P = 7.40E-06 | ||||||
2068 | Apolipoprotein A-1; ApoA1; APOA1-HUMAN; P02647/NP_000030.1 | -2.25 | -2.24 | -2.24 | 30,777.83 | 5.56 |
P = 0.00035 |
Charge-based distribution of differentially charged ApoE isoforms distinguishes AD from non-AD samples
Plasma ApoE charge-isoform distribution may be able to predict AD
Sample number | APOE genotype | Disease state from clinic | Disease state predicted |
---|---|---|---|
1 | 3/4 | No dementia | No disease |
2 | 3/4 | No dementia | No disease |
3 | 3/4 | No dementia | No disease |
4 | 3/3 | No dementia | No disease |
5 | 3/3 | No dementia | No disease |
6 | 3/3 | No dementia | No disease |
7 | 3/3 | Dementia | Disease |
8 | 3/3 | Dementia | Disease |
9 | NR | Dementia | Disease |
10 | 3/3 | No dementia | No disease |
11 | 4/4 | No dementia |
Disease
a
|
12 | 4/4 | No dementia |
Disease
a
|
13 | 4/4 | Dementia | Disease |
14 | 3/4 | Dementia | Disease |
15 | 4/4 | Dementia | Disease |
16 | 3/4 | Dementia | Disease |
17 | 3/4 | Dementia | Disease |
18 | 3/4 | Dementia | Disease |