Proteome Survey Reveals Modularity of the Yeast Cell Machinery:Global Landscape of Protein Complexes in the Yeast Saccharomyces cervisiae

摘要

Proteins act often in concert to form protein complexes: successive reactions within one biochemical pathway are organized in one complex, so that the order of reactions is reflected in the composition of that complex. Replication of the genome, transcription, and RNA metabolism, as well as stress responses, energy metabolism, and defense against pathogens, are realized by protein complexes. Yeast was the first eukaryotic organism whose genome was completely sequenced. Its smaller genome (e.g., in comparison to mammals) and its long history in physiological and biochemical investigations make it an important eukaryotic model organism. Different methods elucidate the composition of protein complexes such as coimmuno-precipitation, yeast two-hybrid screens, and phage display. High-throughput methods combined with bioinformatics aim at resolving the complete "inter-actome" of an entire organism. This effort was started some years ago by new tandem affinity purification (TAP) protocols that obtain highly pure complexes. The technique used overexpression of proteins having two affinity tags in a row, protein A and calmodulin-binding protein.