Purifying protein complexes for mass spectrometry: applications to protein translation.
Proteins control and mediate most of the biological activities in the cell. In most cases, proteins either interact with regulatory proteins or function in large molecular assemblies to carryout biological processes. Understanding the functions of individual proteins requires the identification of these interacting proteins. With its speed and sensitivity, mass spectrometry has become the dominant method for identifying components of protein complexes. This article reviews and discusses various approaches to purify protein complexes and analyze the proteins using mass spectrometry. As examples, methods to isolate and analyze protein complexes responsible for the translation of messenger RNAs into polypeptides are described.
Link
Andrew J
AJ
Department of Microbiology and Immunology, Vanderbilt University School of Medicine, Nashville, TN 37232-8575, USA. andrew.link@vanderbilt.edu
Fleischer
Tracey C
TC
Weaver
Connie M
CM
Gerbasi
Vincent R
VR
Jennings
Jennifer L
JL
D002454:Cell Differentiation; D002499:Centrifugation, Density Gradient / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D047468:Immunoprecipitation; D013058:Mass Spectrometry / Q000379:methods*; D008954:Models, Biological; D014176:Protein Biosynthesis; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification*; D040901:Proteomics / Q000379:methods*; D012333:RNA, Messenger / Q000378:metabolism; D012270:Ribosomes / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D012680:Sensitivity and Specificity; D013395:Sucrose / Q000494:pharmacology; D013696:Temperature; D013997:Time Factors
D000939:Epitopes; D011506:Proteins; D012333:RNA, Messenger; D013395:Sucrose