Sequencing of Mammalian Genomes Predicts 30,000 genes
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Sequencing of Mammalian Genomes Predicts 30,000 genes 2001 2002
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Sequencing of Mammalian Genomes Predicts 30,000 genes. 2001. 2002. What is the Proteome. All the proteins expressed in a particular cell or tissue. By definition this will vary by tissue and cell type. Cardiovascular Neuromuscular Islet cell Endothelial cell Muscle cell. - PowerPoint PPT Presentation
Transcript of Sequencing of Mammalian Genomes Predicts 30,000 genes
Sequencing of Mammalian Genomes Predicts 30,000 genes
2001 2002
What is the Proteome
• All the proteins expressed in a particular cell or tissue.
• By definition this will vary by tissue and cell type.
– Cardiovascular– Neuromuscular– Islet cell– Endothelial cell– Muscle cell
Why Study Proteomics?
• Gives a better understanding of the function of gene products.
• Allow for the design of rational drug therapies.
• Provide new and specific markers of disease.
Will Proteomics Provide a Stronger Means to Stratify Disease
Greater Complexity=Greater Specificity
YES
Protein Modifications
PhosphorylationGlycosolationUbiquitinationCleavageLipid etc.
Dictates Function and Intracellular Localization
30,000 Genes How Many Proteins?
30,000 genes
4-6 splice variants
120,000 possible mRNAS
>200 Post-translational Modifications
24,000,000 Possible Protein Isoforms
2,000,000 Predicted Proteins
How Gene and Protein Expression is Regulated or Modified from Transcription to
Post-translation
Synaptojanin 2
Biomarker ID vrs Protein ID
BIOMARKER• CHANGE +/-• IDENTITY ?• SPECIFIC• EASE• BODY FLUID
PROTEIN• INTERACTION• PRESENCE• ISOFORMS• MODIFICATIONS• FUNCTION
Using Proteomics to Identify Biomarkers of Disease
How do You ID Biomarkers
• 2D-PAGE MALDI-TOF MS
• ICAT MALDI-TOF MS• SELDI-TOF MS
Proteomic flow
1D and 2D gels
Proexpress Imager Progest ProMSPropicker
Q Star XL-MS/MS-TOF
Vision StationBiacore
ICAT(Isotope Coded
Affinity Tag)
Deuterium
Biotin Linker Thiol ReactiveGroup
ICAT Provides Relative Levels of
Expression Between Two
Proteins Which is
Reflected by the Ratio of
the Amount of Peptide
Surface Enhanced Laser Desorption Ionization Time of Flight (SELDI-TOF)
ProteinChip
Protein Chip
Different Protein Profiles from Different Chip Surfaces
Laser Capture
Microscopy Allows for the Dissection of Cells Directly
from Sectioned Tissues
LCM of Normal and Cancerous Prostate
Protein Profile from Prostrate Cancer Tissue
Procured by LCM
Do we Need all Three Approaches?• 2D-PAGE MALDI-TOF MS
– Greatest resolving power– Large data base– Labor and time consuming
• ICAT MALDI-TOF MS– Mostly abundant proteins– Needs cysteine residue
• SELDI-TOF MS– Resolves from 8-50 kDa– Small amounts of sample
