“Proteomics is a science that focuses on the study of proteins: their roles, their structures, their localization, their interactions, and other factors.” www.lexicon-biology.com

The “omics” nomenclature… Genomics DNA (Gene) Functional Transcriptomics RNA Proteomics PROTEIN Metabolomics METABOLITE Transcription Translation Enzymatic reaction

Kinds of Proteomics Functional Proteomics Expressional Proteomics The identification of protein functions, activities or interactions at a global or organismwide scale. Expressional Proteomics The analysis of global or organismwide changes in protein expression. Structural Proteomics The high throughput, or high volume expression and structure determination of proteins by Xray, NMR or computer based methods.

Components of Expressional Proteomics Protein Separation Mass Spectroscopy Bioinformatics

Applications: Biology and disease research Molecular event description (signaling molecules, biomarkers) Finding disease mechanisms and drug targets

Roles of Proteins Proteins are the instruments through which the genetic potential of an organism are expressed = active biological agents in cells Proteins are involved in almost all cellular processes and fulfill many functions *enzyme catalysis, transport, mechanical support, organelle constituents, storage reserves, metabolic control, protection mechanisms, toxins, and osmotic pressure

Why Proteomics? Whole Genome Sequence –complete, but does not show how proteins function or biological processes occur Post-translational modification *proteins sometimes chemically modified or regulated after synthesis Proteins fold into specific 3-D structures which determine function Gain insight into alternative splicing Aids in genome annotation

The Human Proteome Initiative. (2007) http://ca. expasy The Human Proteome Initiative. (2007) http://ca.expasy.org/sprot/hpi/hpi_desc.html Retrieved March 24, 2009.

Work flow and Technologies Proteomics Work flow and Technologies 2-dimensional gel based technologies Mass spectrometry for protein identification and quantification

11/17/2018

2-D Gel Electrophoresis 1st dimension: Isoelectric focusing (IEF) separate proteins based on their isoelectric point (pI) by using immobilized pH gradient (IPG) gel strips. 2nd dimension: SDS PAGE separate proteins based on their molecular mass. run perpendicular to Isoelectric focusing. 11/17/2018

11/17/2018

2-dimensional gel based technologies 11/17/2018