Abstract

We are witnessing tremendous advances in our understanding of the organization of life. Complete genomes are being deciphered with ever increasing speed and accuracy, thereby setting the stage for addressing the entire gene product repertoire of cells, towards understanding whole biological systems. Advances in bioinformatics and mass spectrometric techniques have revealed the multitude of interactions present in the proteome. Multiprotein complexes are emerging as a paramount cornerstone of biological activity, as many proteins appear to participate, stably or transiently, in large multisubunit assemblies. Analysis of the architecture of these assemblies and their manifold interactions is imperative for understanding their function at the molecular level. Structural genomics efforts have fostered the development of many technologies towards achieving the throughput required for studying system-wide single proteins and small interaction motifs at high resolution. The present shift in focus towards large multiprotein complexes, in particular in eukaryotes, now calls for a likewise concerted effort to develop and provide new technologies that are urgently required to produce in quality and quantity the plethora of multiprotein assemblies that form the complexome, and to routinely study their structure and function at the molecular level. Current efforts towards this objective are summarized and reviewed in this contribution.

Highlights

  • Protein-protein interactions (PPIs) are intrinsic to virtually every essential process in the cell

  • By modifying the tandem affinity purification (TAP) tag to include green fluorescent protein (GFP) followed by the Z-domain of protein G instead of protein A, and by replacing the calmodulin-binding peptide (CBP)-tag with streptavidin peptide, this study revealed intact complexes involved in C. elegans kinetochore formation

  • The transfer vectors that we developed for MultiBac contained elements that made it straight forward to arrange into multigene expression cassettes several to many expression units containing open reading frames (ORFs) encoding for example for members of a protein complex of choice

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Summary

Getting a Grip on Complexes

Yan Nie#, Cristina Viola#, Christoph Bieniossek, Simon Trowitzsch, Lakshmi Sumitra Vijayachandran, Maxime Chaillet, Frederic Garzoni and Imre Berger*. European Molecular Biology Laboratory (EMBL), Grenoble Outstation and Unit of Virus Host-Cell Interactions (UVHCI), UJF-EMBL-CNRS, UMR 5233, 6 rue Jules Horowitz, 38042 Grenoble CEDEX 9, France

INTRODUCTION
DECIPHERING THE INTERACTOME
Purification from Native Source
Computational Approaches
Mass Spectrometry
IMPACT OF STRUCTURAL GENOMICS
ACEMBLING MULTIPROTEIN COMPLEXES
Findings
STRUCTURAL COMPLEXOMICS?
Full Text
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