A PRIMER OF GENOME SCIENCE.ppt

A PRIMER OF GENOME SCIENCE Chapter 4 Proteomics and Functional Genomics Hao Sun Index Introduction Qualitative methodologies 2D electrophoresis Mass spectrometry(MS) Quantitative methodologies X-ray crystallography Nuclear magnetic resonance(NMR) Spectroscopy The Nobel Prize in Chemistry 2002 Introduction 2D electrophoresis 2D electrophoresis Mass spectrometry(MS)A powerful technique for measuring the mass of proteins Mass spectrometry(MS) Other qualitative methods for protein X-ray crystallography X-ray crystallography Nuclear magnetic resonance(NMR) Spectroscopy Protein Structure Determination * * ??????????????????????????????? ??????????????????????????????? ??????????????????????????????? John B. Fenn Koichi Tanaka Kurt Wüthrich USA Japan Switzerland ????????????????????? for the development of methods for identification and structure analyses of biological macromolecules for their development of soft desorption ionisation methods for mass spectrometric analyses of biological macromolecules“ for his development of nuclear magnetic resonance spectroscopy for determining the three-dimensional structure of biological macromolecules in solution“ Copyright? 2003 The Nobel Foundation What is Proteomics? Proteomics is the study of the structure and expression of proteins and of the interactions between proteins. Focus on how data on the expression and identity of proteins in cell is obtained. Include several methods that are used to study the structure of proteins. Key features: Predicted Protein domains from sequence alignment. Proteins are separated in two sequential steps: first by their charge and then by their mass. Isoelectric focusing (IEF) can resolve proteins that differ by only one charge unit. Charged proteins will migrate through the PH gradient until they reach their isoelectric point. When an electric field is imposed, the proteins will migrate from the IEF gel into the SDS slab gel and then separate according to their mass. For