Introduction
Materials and methods
Materials
Human anterior temporal lobe samples
Sample ID | Case | Age (years) | Gender | PMI (h) | pH values | Type of SCZ | Duration of disease (years) | Duration of medication (years) | atyptyp | CPE last dosis | CPE last 10 years | Cause of death | DSM IV | Age at onset | Last medication | Cigarettes | Alcohol | Hosp | ECT |
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
01/00 | SCZ | 51 | M | 12 | 6.7 | Residual, Chronic Paranoid episodes | 28 | 25 | 2 | 450 | 1.8 | Heart infarction | 295.6 | 23 | Clozapine 500 mg | 30/day | No | 17 | No |
35/00 | SCZ | 64 | F | 23 | 6.6 | Residual, Chronic Paranoid episodes | 41 | 40 | 2 | 54.5 | 4.6 | Heart infarction | 295.6 | 24 | Zotepine 150 mg Olanzapine 10 mg | 20/day | No | 5 | Yes |
36/02 | SCZ | 73 | M | 20 | 6.9 | Residual, Chronic Paranoid episodes | 43 | 40 | 1 | 507.4 | 1.7 | Heart infarction | 295.6 | 30 | Perphenazine 32 mg Promethazine 150 mg | 30/day | No | 33 | No |
83/01 | SCZ | 71 | M | 28 | 6.5 | Residual, Chronic Paranoid episodes | 40 | 35 | 1 | 782.4 | 10 | Heart infarction | 295.6 | 30 | Haloperidol 32 mg Pipamperone 40 mg | 40/day | No | 12 | No |
50/01 | SCZ | 81 | M | 4 | 6.8 | Residual, Chronic Paranoid episodes | 62 | 50 | 1 | 92.8 | 1.4 | Cor pulmonale, heart insufficiency | 295.6 | 19 | Haloperidol 4 mg Prothypendyl 80 mg | 20/day | No | 48 | No |
43/01 | Control | 91 | F | 16 | 6.8 | Cardio-pulmonary insuffiency | No | No | |||||||||||
57/02 | Control | 53 | M | 18 | 7.1 | Heart infarction | No | No | |||||||||||
61/01 | Control | 66 | M | 16 | 6.9 | Heart infarction | No | No | |||||||||||
72/02 | Control | 79 | M | 24 | 6.4 | Heart infarction | No | No |
Sample preparation
ICPL labeling
Digestion of labeled proteins and fractionation of peptides by isoelectric focusing
Fractionation of peptides by nano high performance liquid chromatography
Mass spectrometry
Identification of proteins
ICPL quantitative analysis
Determination of regulated proteins in SCZ samples
Results
Protein regulation in schizophrenic anterior temporal lobe
Biological process | Regulation in SCZ | Protein name | Gene name | Accession | H/L ratio | pI (th) | MW (th) | Chr loci | Id. pept | MASCOT score |
---|---|---|---|---|---|---|---|---|---|---|
Transport | ↑ | Plasma membrane calcium-transporting ATPase 4 | PMCA-4 | P23634 | 1.59 | 6.19 | 137920.16 | 1q25-q32 | 3 | 76.49 |
↓ | Hemoglobin subunit beta | HBB | P68871 | 0.57 | 6.81 | 15867.22 | 11p15.4-p15.5 | 2 | 44.1 | |
Signal transduction Cell communication | ↑ | Ezrin-radixin-moesin-binding phosphoprotein 50 | EBP50 | O14745 | 1.58 | 5.55 | 38737.24 | 17q25.2 | 2 | 50.76 |
↑ | Prohibitin | PHB | Q6FHP5 | 1.57 | 5.57 | 29832.16 | 17q21 | 2 | 62.38 | |
↑ | Phosphatidylethanolamine-binding protein 1 | PEBP1 | P30086 | 2.86 | 7.43 | 20925.59 | 12q24.23 | 3 | 44.07 | |
↓ | 14-3-3 protein eta | YWHAH | Q04917* | 0.49 | 4.76 | 28087.53 | 22q12-q13 | 2 | 65.33 | |
↓ | 14-3-3 protein gamma | YWHAG | P61981 | 0.46 | 4.8 | 28171.4 | 7q11.23 | 4 | 112.84 | |
↓ | 14-3-3 protein zeta/delta | YWHAZ | P63104 | 0.52 | 4.73 | 27745.1 | 2p25.2-p25.1 | 4 | 58.22 | |
↓ | Calcineurin or serine/threonine-protein phosphatase 2B catalytic subunit alpha isoform | PPP3CA or CALN | Q08209 | 0.60 | 5.58 | 58687.85 | 4q21-q24 | 3 | 64.34 | |
↓ | Visinin-like protein 1 | VILIP-1 | P62763 | 0.51 | 5.01 | 22011.14 | 2p24.3 | 3 | 91.89 | |
↓ | Signal-regulatory protein beta-1 | SIRPB1 | O00241 | 0.56 | 6.06 | 43255.18 | 20p13 | 4 | 118.36 | |
↓ | Calmodulin | CALM | P62158 | 0.20 | 4.09 | 16706.39 | 14q24-q31 | 8 | 131.36 | |
Metabolism; energy pathways | ↑ | Hexokinase brain form | HK1 | P19367 | 1.84 | 6.44 | 102484.91 | 10q22 | 2 | 43.49 |
↓ | NADH-ubiquinone oxidoreductase 13 kDa-A subunit | NDUFS6 | O75380 | 0.58 | 8.58 | 13711.61 | 5p15.33 | 3 | 92.52 | |
↓ | NADH-ubiquinone oxidoreductase SGDH subunit | NDUFB5 | O43674 | 0.48 | 9.62 | 21750.27 | 3q26.33 | 2 | 51.45 | |
↓ | NADH-ubiquinone oxidoreductase 30 kDa subunit | NDUFS3 | O75489 | 0.53 | 6.98 | 30241.53 | 11p11.11 | 4 | 81.08 | |
↓ | NADH dehydrogenase (ubiquinone) flavoprotein 2 | NDUFV2 | Q6IB76 | 0.48 | 8.22 | 27349.47 | 18p11.31-p11.2 | 2 | 112.02 | |
↓ | Carbonic anhydrase 2 | CA2 | P00918 | 0.50 | 6.86 | 29114.86 | 8q22 | 2 | 74.22 | |
↓ | Fructose bisphosphate; aldolase C | ALDOC | Q6FH94 | 0.56 | 6.41 | 39455.87 | 17cen-q12 | 5 | 79.21 | |
↓ | 2′,3′-Cyclic-nucleotide 3′-phosphodiesterase | CNP | P09543 | 0.51 | 9.17 | 47578.63 | 17q21 | 13 | 87.29 | |
Cell growth and/or maintenance | ↑ | Tubulin, beta polypeptide | TUBB | Q9BVA1* | 1.46 | 4.78 | 49953.06 |
6p25
| 3 | 83.45 |
↓ | Aggrecan core protein | AGC | P16112* | 0.50 | 4.1 | 250193.19 | 15q26.1 | 2 | 56.55 | |
↓ | Spectrin alpha chain; Fodrin | SPTAN1 | Q13813 | 0.43 | 5.22 | 284539.36 | 9q33-q34 | 2 | 55.93 | |
↓ | Glial fibrillary acidic protein, astrocyte | GFAP | P14136 | 0.59 | 5.42 | 49880.21 | 17q21 | 16 | 103.48 | |
↓ | Neurofilament triplet M protein | NEFM | P07197* | 0.46 | 4.9 | 102316.8 | 8p21 | 3 | 68.45 | |
↓ | Hyaluronan and proteoglycan link protein 2 | HAPLN2 | Q5T3J1* | 0.16 | 8.37 | 20827.82 | 1q23.1 | 3 | 82.95 | |
↓ | Neurofilament triplet L protein | NEFL | P07196* | 0.32 | 4.64 | 61385.41 | 8p21 | 2 | 70.24 | |
↓ | Clathrin light chain B | CLTB | P09497* | 0.50 | 4.57 | 25190.49 | 5q35 | 3 | 74.98 | |
Protein metabolism | ↓ | Ubiquitin-conjugating enzyme E2N | UBE2 N | P61088 | 0.32 | 6.13 | 17137.82 | 12q21.33 | 2 | 82.01 |
↓ | Eukaryotic initiation factor 4A-II | EIF4A2 | Q14240 | 0.52 | 5.33 | 46402.27 | 3q28 | 2 | 46.03 | |
↓ | 60S ribosomal protein L13 | RPL13 | P26373 | 0.46 | 11.65 | 24130.28 | 16q24.3 | 2 | 62.73 | |
↓ | 40S ribosomal protein S10 | RPS10 | P46783* | 0.53 | 10.15 | 18897.77 | 6p21.31 | 2 | 54.19 | |
Nucleic acid metabolism | ↓ | Histone H3.1 | HIST1H3E | P68431* | 0.31 | 11.13 | 15272.89 | 6p21.3 | 2 | 58.35 |
↓ | Ribonuclease/angiogenin inhibitor 1 | RNH1 | P13489 | 0.59 | 4.71 | 49842.28 | 11p15.5 | 2 | 82.56 | |
Immune response Neurogenesis Synaptic Transmission Oligodendrocyte metabolism | ↓ | Myelin oligodendrocyte glycoprotein | MOG | Q5SSB7* | 0.43 | 8.29 | 23235.99 | 6p22.1 | 3 | 88.54 |
↓ | Myelin basic protein | MBP | Q65ZS4 | 0.39 | 11.35 | 22316.06 | 18q22-qter | 7 | 78.43 | |
↓ | Ermin (myelinating oligodendrocyte-specific protein) | ERMN | Q8TAM6 | 0.31 | 4.75 | 32782.95 | 2q24.1 | 2 | 61.42 |
Functional classification of regulated proteins
Discussion
ICPL methodology for proteomic analysis
Shotgun proteomics confirms the regulation of previously identified SCZ-related proteins
Oligodendrocyte-myelin regulated proteins
Regulation of Ca2+ homeostasis
Energy metabolism
Other regulated proteins
Gene symbol | Product name | Type of analysis | Tissue | Described as regulated by |
---|---|---|---|---|
MOG | Myelin oligodendrocyte glycoprotein | 1. Microarray and qPCR 2. Microarray 3. Microarray and qPCR | 1. Pre-frontal cortex 2. Several brain region 3. Pre-frontal cortex | 1. Tkachev et al. 2003
2. Katsel et al. 2005a
3. Arion et al. 2007
|
MBP | Myelin basic protein | 1. Immunoassay 2. Microarray and qPCR | 1. Anterior frontal cortex 2. Pre-frontal cortex | 1. Honer et al. 1999
2. Tkachev et al. 2003
|
GFAP | Glial fibrillary acidic protein | 1. Proteomics 2. In situ hybridization 3. Microarray and qPCR | 1. Frontal cortex 2. Cingulate cortex 3. Pre-frontal cortex | 1. Johnston-Wilson et al. 2000
2. Webster et al. 2005
3. Tkachev et al. 2003
|
CNP | 2′,3′-Cyclic-nucleotide 3′-phosphodiesterase | 1. Microarray 2. Microarray and qPCR 3. Microarray 4. Microarray and proteomics 5. Microarray 6. Microarrays 7. qPCR | 1. Cortical tissues 2. Pre-frontal cortex 3. Temporal gyrus 4. Pre-frontal cortex 5. Several brain region 6. Anterior cingulate cortex and hippocampus 7. Anterior cingulate cortex | 1. Hakak et al. 2001
2. Tkachev et al. 2003
3. Aston et al. 2004
4. Prabakaran et al. 2004
5. Katsel et al. 2005a
6. Dracheva et al. 2006. 7. McCullumsmith et al. 2007
|
PPP3CA or CALN | Calcineurin | 1. Microarray 2. Microarray | 1. Cortical tissues 2. Hippocampus | 1. Hakak et al. 2001
2. Eastwood et al. 2005
|
SPTAN1 | Spectrin alpha chain (fodrin) | Western blot | Left superior temporal cortices | Kitamura et al. 1998
|
14-3-3 eta (YWHAH) | Protein kinase C inhibitor protein 1 (eta) | 1. In situ hybridization 2. Microarray | 1. Pre-frontal cortex 2. Cerebellum | 1. Middleton et al. 2005
2. Vawter et al. 2001
|
14-3-3 zeta/delta (YWHAZ) | Protein kinase C inhibitor protein 1 (zeta/delta) | 1. In situ hybridization 2. Proteomics | 1. Pre-frontal cortex 2. Corpus callosum | 1. Middleton et al. 2005
2. Sivagnanasundaram et al. 2007
|
14-3-3 gamma (YWHAG) | Protein kinase C inhibitor protein 1 (gamma) | 1. In situ hybridization 2. Proteomics | 1. Pre-frontal cortex 2. Corpus callosum | 1. Middleton et al. 2005
2. Sivagnanasundaram et al. 2007
|
ALDOC | Fructose bisphosphate aldolase C | 1. Proteomics 2. Microarray and proteomics 3. Proteomics 4. Immunoassay* | 1. Anterior cingulate cortex 2. Pre-frontal cortex 3. Frontal cortex 4. Human CSF | 1. Clark et al. 2006
2. Prabakaran et al. 2004
3. Johnston-Wilson et al. 2000
4. Willson et al. 1980
|
HIST1H3A | Histone H3.1 | Immunoblotting, immunohistochemical, microarray and qPCR | Pre-frontal cortex | Akbarian et al. 2005
|
CA2 | Carbonic anhydrase 2 | 1. Proteomics 2. Proteomics | 1. Frontal cortex 2. Anterior cingulate cortex | 1. Johnston-Wilson et al. 2000
2. Beasley et al. 2006
|
RPS10 | 40S ribosomal protein—family S10 | Microarray | Pre-frontal cortex | Vawter et al. 2002
|
EBP50 | Ezrin-radixin-moesin-binding phosphoprotein 50 | Microarray | Peripheral blood lymphocytes | Bowden et al. 2006
|
TUBB | Tubulin beta polypeptide | 1. In situ hybridization 2. Proteomics | 1. Frontal cortex 2. Corpus callosum | 1. Virgo et al. 1995
2. Sivagnanasundaram et al. 2007
|
NEFL | Neurofilament triplet L protein | Proteomics | Corpus callosum | Sivagnanasundaram et al. 2007
|
NEFM | Neurofilament triplet M protein | Proteomics | Corpus callosum | Sivagnanasundaram et al. 2007
|