Proteomics - Technologies, Markets and Companies

0. Executive Summary .... 15

1. Basics of Proteomics... 17

Introduction...... 17

History ............... 17

Nucleic acids, genes and proteins ........... 18

Genome..........18

DNA ................19

RNA................19

MicroRNAs...19

Decoding of mRNA by the ribosome.....20

Genes.............20

Alternative splicing ...............20

Transcription ....21

Gene regulation 22

Gene expression 22

Chromatin.......23

Proteins ...........23

Spliceosome 23

Functions of proteins .......24

Inter-relationship of protein, mRNA and DNA...........24

Proteomics........ 25

Mitochondrial proteome ........26

S-nitrosoproteins in mitochondria........27

Proteomics and genomics ..... 27

Classification of proteomics ...29

Levels of functional genomics and various "omics" ........30

Glycoproteomics 30

Transcriptomics 30

Metabolomics ...31

Cytomics.........31

Phenomics ......... 31

Proteomics and systems biology ............. 32

2. Proteomic Technologies................. 33

Key technologies driving proteomics....... 33

Sample preparation............... 34

New trends in sample preparation ............34

Protein separation technologies.............. 35

High resolution 2D gel electrophoresis ......35

Variations of 2D gel technology................35

Limitations of 2DGE and measures to overcome these ...36

1-D sodium dodecyl sulfate (SDS) PAGE ...36

Capillary electrophoresis systems.............36

Head column stacking capillary zone electrophoresis .....37

Removal of albumin and IgG..37

Companies with protein separation technologies ...........37

Protein detection  39

Protein identification and characterization ................ 39

Mass spectrometry (MS) .......39

Companies involved in mass spectrometry ..............40

Electrospray ionization .....41

Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry...42

Cryogenic MALDI- Fourier Transform Mass Spectrometry ............43

HUPO Gold MS Protein Standard..........43

High performance liquid chromatography ..44

Multidimensional protein identification technology (MudPIT) .............44

Peptide mass fingerprinting ...44

Combination of protein separation technologies with mass spectrometry...............45

Combining capillary electrophoresis with mass spectrometry .......45

2D PAGE and mass spectrometry ........45

Quantification of low abundance proteins ................45

Antibodies and proteomics ....46

Labeling and detection of proteins ............46

Fluorescent labeling of proteins in living cells ...........46

Combination of microspheres with fluorescence .......47

Self-labeling protein tags .47

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Analysis of peptides..............47

Differential Peptide Display.................48

Peptide analyses using NanoLC-MS......49

Protein sequencing ...............50

Functional proteomics: technologies for studying protein function................. 50

RNA-Protein fusions.............. 50

Designed repeat proteins...... 51

Application of nanbiotechnology to proteomics.......... 51

Nanoproteomics51

Protein nanocrystallography ...............52

Single-molecule mass spectrometry using a nanopore ................52

Nanoelectrospray ionization................52

Nanoparticle barcodes ..........53

Biobarcode assay for proteins .............53

Resonance Light Scattering technology .....55

Nanoscale protein analysis ....55

Nanobiotechnology for discovery of protein biomarkers in the blood ..55

Study of single membrane proteins at subnanometer resolution........56

Nanotube-vesicle networks for study of membrane proteins .............56

Qdot-nanocrystals................57

Nanotube electronic biosensor .................57

A nanoscale mechanism for protein engineering............57

Protein expression profiling .. 58

Protein function studies ........ 58

Transcriptionally Active PCR ..58

Cell-based protein function assays ...........59

Protein-protein interactions ...59

Yeast two-hybrid system..61

Membrane one-hybrid method ............62

Protein affinity chromatography ..........62

Phage display .................62

Fluorescence Resonance Energy Transfer ................63

Bioluminescence Resonance Energy Transfer ...........63

Detection Enhanced Ubiquitin Split Protein Sensor technology......64

Protein-fragment complementation system..............64

In vivo study of protein-protein interactions ............64

Computational prediction of interactions .................65

Interactome 65

Protein-protein interactions and drug discovery........66

Companies with technologies for protein-protein interaction studies ................66

Protein-DNA interaction ........67

Determination of protein structure ...........67

X-Ray crystallography...........68

Nuclear magnetic resonance..69

Electron spin resonance ...69

Prediction of protein structure70

Protein tomography ..............70

Prediction of protein function .71

Three-dimensional proteomics for determination of function ........71

An algorithm for genome-wide prediction of protein function .......72

Monitoring protein function by expression profiling ...72

Isotope-coded affinity tag peptide labeling................ 72

Differential Proteomic Panning ............... 73

Cell map proteomics.............. 73

Topological proteomics ......... 73

Organelle or subcellular proteomics........ 74

Nucleolar proteomics ............ 75

Glycoproteomic technologies  75

High-sensitivity glycoprotein analysis........75

Fluorescent in vivo imaging of glycoproteins.................75

Integrated approaches for protein characterization... 76

Imaging mass spectrometry ..76

The protein microscope.........77

Automation and robotics in proteomics... 77

Laser capture microdissection ................ 78

Microdissection techniques used for proteomics ............78

Uses of LCM in combination with proteomic technologies78

Concluding remarks about applications of proteomic technologies................. 79

3. Protein biochip technology ............ 81

Introduction...... 81

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Types of protein biochips ...... 82

ProteinChip ......82

Applications and advantages of ProteinChip .............83

ProteinChip Biomarker System............83

Matrix-free ProteinChip Array..............84

Lab-on-a-chip for protein analysis ............84

Protein biochips based on fluorescence planar wave guide technology85

Application of microfluidic biochips in proteomics ..........85

Aptamer-based protein biochip ................86

High-density protein microarrays .............86

Protein biochips with a functional continuum.................87

Tissue microarray technology for proteomics ............ 87

Protein biochips for high-throughput expression....... 88

Antibody microarrays............89

Protein biochips in molecular diagnostics  89

A force-based protein biochip 90

L1 chip and lipid immobilization ...............90

Multiplexed Protein Profiling on Microarrays90

Live cell microarrays ............. 91

ProteinArray Workstation ..... 91

Proteome arrays  92

The Yeast ProtoArray............92

ProtoArrayÔ Human Protein Microarray.....92

TRINECTIN proteome chip.....93

Peptide arrays.... 93

Surface plasmon resonance technology .. 94

Biacore's SPR ...94

FLEX CHIP........94

Combination of surface plasmon resonance and MALDI-TOF .............95

Protein chips/microarrays using nanotechnology...... 95

Nanoparticle protein chip ......95

Protein nanobiochip ..............96

Protein nanoarrays ...............96

Self-assembling protein nanoarrays..........96

Companies involved in protein biochip/microarray technology.... 97

4. Bioinformatics in Relation to Proteomics ....... 101

Introduction.... 101

Bioinformatic tools for proteomics........ 101

Testing of SELDI-TOF MS Proteomic Data 101

BioImagine’s ProteinMine .... 102

Bioinformatics for pharmaceutical applications of proteomics ... 102

In silico search of drug targets by Biopendium ............ 102

Compugen's LEADS ............ 103

DrugScore...... 103

Proteochemometric modeling................. 103

Integration of genomic and proteomic data ............. 104

Proteomic databases: creation and analysis ............ 105

Introduction ... 105

Proteomic databases .......... 105

Human Proteomics Initiative 106

Protein Warehouse Database ................. 106

GenProtEC...... 106

Human Protein Index.......... 107

Protein Data Bank .............. 107

Universal Protein Resource .. 107

Protein interaction databases................. 108

Biomolecular Interaction Network Database................ 109

Proteome maps 109

The Protein Atlas................ 109

ENCODE......... 110

Metabolomics databases ..... 110

Functional Genomics Consortium............ 110

Application of bioinformatics in functional proteomics .............. 110

Us of bioinformatics in protein sequencing .............. 111

Bottom-up protein sequencing ............... 111

Top-down protein sequencing ................ 112

Protein structural database approach to drug design................. 112

Bioinformatics for high-throughput proteomics ....... 113

Companies with bioinformatic tools for proteomics . 114

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5. Research in Proteomics ............... 117

Introduction.... 117

Basic biological research..... 117

Identification of novel human genes by comparative proteomics ..... 118

Study of relationship between genes and proteins ....... 118

Protein knockout ................ 118

Antisense approach and proteomics... 118

RNAi and protein knockout ............... 118

Total knockout of cellular proteins ..... 119

Ribozymes and proteomics .. 119

Single-molecule photon stamping spectroscopy .......... 119

Functional genomics by mass spectrometry................ 120

Stable-isotope-dilution tandem mass spectrometry ..... 120

Single nucleotide polymorphism determination by TOF-MS ............. 120

Application of proteomic technologies in systems biology ............... 121

Signaling pathways and proteomics ... 121

Kinomics ... 121

Combinatorial RNAi for quantitative protein network analysis..... 122

Proteomics in neuroscience research ...... 122

Stem cell proteomics .......... 123

Proteomic studies of mesenchymal stem cells ........ 123

Proteomic profiling of neural stem cells ................. 123

Proteome Biology of Stem Cells Initiative .............. 124

Proteomic analysis of the cell cycle......... 124

Nitric oxide and proteomics . 124

A proteomic method for identification of cysteine S-nitrosylation sites ........... 125

Study of the nitroproteome .............. 125

Study of the phosphoproteome .............. 125

Study of the mitochondrial proteome...... 125

Proteomic technologies for study of mitochondrial proteomics.... 126

Cryptome....... 126

Study of protein transport in health and disease ......... 127

Proteomics research in the academic sector................ 127

Vanderbilt University's Center for Proteomics and Drug Actions.. 129

ProteomeBinders initiative................ 129

6. Pharmaceutical Applications of Proteomics ... 131

Current drug discovery process and its limitations ....... 131

Genomics-based drug discovery............. 132

Metabolomics technologies for drug discovery............. 132

Role of metabonomics in drug discovery . 133

Proteins and drug action ..... 133

Transcription-aided drug design ............. 134

Role of proteomic technologies in drug discovery ........ 134

Protein-expression mapping by 2DGE  135

Liquid chromatography-based drug discovery ........ 135

Role of MALDI mass spectrometry in drug discovery ................. 135

Oxford Genome Anatomy Project ...... 137

Proteins as drug targets...... 138

Ligands to capture the purine binding proteome.......... 138

Chemical probes to interrogate key protein families for drug discovery ............... 138

CellCarta® proteomics platform............. 139

ZeptoMARKÔ protein profiling system..... 139

Identification of protein kinases as drug targets .......... 140

G-protein coupled receptors as drug targets ............... 140

Methods of study of GPCRs............... 141

Companies involved in GPCR-based drug discovery  141

GPCR localization database............... 142

Targeting mTOR signaling pathway......... 142

Serine hydrolases as drug targets .......... 143

Matrix metalloproteases as drug targets.. 144

Integrated bioinformatic analysis of proteomics data for drug discovery.............. 144

Drug design based on structural proteomics ............... 144

Protein crystallography for determining 3D structure of proteins  145

Automated 3D protein modeling........ 145

Companies involved in structure-based drug-design ................. 145

Integration of genomics and proteomics for drug discovery ............ 146

Ligand-receptor binding ...... 147

Aptamer protein binding ..... 147

Systematic Evolution of Ligands by Exponential Enrichment....... 147

Aptamers and high-throughput screening .............. 148

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Nucleic Acid Biotools...... 149

Aptamer beacons .......... 149

Peptide aptamers .......... 149

Riboreporters.. 150

Target identification and validation......... 150

Application of mass spectrometry for target identification.......... 151

Gene knockout and gene suppression for validating protein targets ............... 151

Laser-mediated protein knockout for target validation .............. 151

Integrated proteomics for drug discovery .............. 152

High-throughput proteomics  152

Companies involved in high-throughput proteomics 153

Drug discovery through protein-protein interaction studies............. 153

Protein-protein interaction as basis for drug target identification  154

Protein-PCNA interaction as basis for drug design... 154

Two-hybrid protein interaction technology for target identification................. 155

Biosensors for detection of small molecule-protein interactions .. 155

Protein-protein interaction maps ............ 156

ProNet (Myriad Genetics) ................. 156

Hybrigenics' maps of protein-protein interactions ... 156

CellZome's functional map of protein-protein interactions.......... 157

Mapping of protein-protein interactions by mass spectrometry... 157

Protein interaction map of Drosophila melanogaster ................. 158

Protein-interaction map of Wellcome Trust Sanger Institute....... 158

Inhibition of protein-protein interactions by peptide aptamers ........ 158

Selective disruption of proteins by small molecules...... 159

Post-genomic combinatorial biology approach............. 159

Differential p