国家人类基因组北方研究中心课件.pptVIP

Protein Complex and Protein-protein Interaction;Central dogma: the story of life;Proteins function in association with other proteins or biomolecules, but not in isolation;Introduction to Proteomics;Goals of Proteomics;Types of Proteomics;Approaches;Blue Native PAGE;Blue Native PAGE;Blue Native PAGE;Blue Native PAGE of chloroplast thylakoid membranes;Blue Native PAGE of chloroplast thylakoid membranes;Blue Native PAGE of multiprotein complex from whole cellular lysate;Identification and analysis of distinct proteasomes by WCL 2D BN/SDS;Visualization of MPCs on a 2D WCL BN/SDS gel;Far Western;Max: functional cloning of a Myc-binding protein;Association of Rb with HIP1;GST Pulldown;Interactions of Cellular Polypeptides with the Cytoplasmic Domain of the Mouse Fas Antigen;GST-mFas fusion proteins;GST-mFas-associated polypeptides from 32S-labeled HeLa, L929, and Jurkat cell lysates;GST-mFas-associated polypeptides are stable to high salt concentrations;Association is blocked by preincubation with a polyclonal antibody against GST-mFas;Differential association with mutant forms of GST-mFas;Schematic representation of the mouse Fas antigen and its binding proteins;Epitope tagging;Co-Immunoprecipitation;Co-Immunoprecipitation;False positive and control;Reduction of nonspecific protein background; Binding of pVHL to Elongin B and C;Identification of VHL-associated proteins;Mapping the p14 and p18 binding site on pVHL;the binding site for Elongin B and C in pVHL;Binding of pVHL to Elongin B and Elongin C in vivo;TAP: tandem affinity purification;Sequence and structure of the TAP tag;Overview of the TAP procedure;Schematic representation of the split TAP tag strategy;Schematic representation of the substraction strategy;Protein composition of TAP-purified U1 snRNP;Step-by-step analysis of the TAP strategy;TAP in higher eucaryotes;Strengths and weaknesses of commonly used affinity approaches for the retrieval of protein complexes;FRET: fluorescence resonance energy transfer;