Abstract
Cellular function relies on protein complexes that work as nano-machines. The structure and function of protein complexes is an outcome of the specific combination of protein subunits, or modules, within the complex. A major focus of molecular biology is thus to understand how protein subunits assemble to form complexes with distinct biological function. To this end, in vitro reconstitution of complexes from individual subunits to study their assembly, structure and activity is of central importance. With purified individual subunits and sub-modules at hand one can systematically dissect the hierarchical assembly of larger complexes using direct protein-protein interaction assays. Furthermore, activity assays can be carried out with individual subunits or smaller sub-complexes and compared to those of the fully assembled complex to precisely map functional sites and provide a molecular basis for in vivo observations. In this chapter we review methods for protein complex assembly from individual subunits and provide examples of advantages and potential pitfalls to this approach.
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References
Bhogaraju S, Engel BD, Lorentzen E (2013) Intraflagellar transport complex structure and cargo interactions. Cilia 2(1):10
Taschner M, Bhogaraju S, Lorentzen E (2012) Architecture and function of IFT complex proteins in ciliogenesis. Differentiation 83(2):S12–S22
Rabut G, Lénárt P, Ellenberg J (2004) Dynamics of nuclear pore complex organization through the cell cycle. Curr Opin Cell Biol 16(3):314–321
Lorentzen E, Basquin J, Conti E (2008) Structural organization of the RNA-degrading exosome. Curr Opin Struct Biol 18(6):709–713
Aloy P, Russell RB (2004) Ten thousand interactions for the molecular biologist. Nat Biotechnol 22(10):1317–1321
Brodersen DE, Nissen P (2005) The social life of ribosomal proteins. FEBS J 272(9):2098–2108
Lorentzen E, Conti E (2005) Structural basis of 3′ end RNA recognition and exoribonucleolytic cleavage by an exosome RNase PH core. Mol Cell 20(3):473–481
Lorentzen E, Conti E (2006) The exosome and the proteasome: nano-compartments for degradation. Cell 125(4):651–654
Basler M, Kirk CJ, Groettrup M (2013) The immunoproteasome in antigen processing and other immunological functions. Curr Opin Immunol 25(1):74–80
Hartwell LH, Hopfield JJ, Leibler S, Murray AW (1999) From molecular to modular cell biology. Nature 402(supp):C47–C52
Hershko A, Ciechanover A (1998) The ubiquitin system. Annu Rev Biochem 67(1):425–479
Peters J-M (2006) The anaphase promoting complex/cyclosome: a machine designed to destroy. Nat Rev Mol Cell Biol 7(9):644–656
Taschner M, Kotsis F, Braeuer P, Kuehn EW, Lorentzen E (2014) Crystal structures of IFT70/52 and IFT52/46 provide insight into intraflagellar transport B core complex assembly. J Cell Biol 207(2):269–282
Frazão C, McVey CE, Amblar M, Barbas A, Vonrhein C, Arraiano CM, Carrondo MA (2006) Unravelling the dynamics of RNA degradation by ribonuclease II and its RNA-bound complex. Nature 443(7107):110–114
Taschner M, Bhogaraju S, Vetter M, Morawetz M, Lorentzen E (2011) Biochemical mapping of interactions within the Intraflagellar Transport (IFT) B core complex. J Biol Chem 286(30):26344–26352
Basquin J, Roudko VV, Rode M, Basquin C, Séraphin B, Conti E (2012) Architecture of the nuclease module of the yeast Ccr4-Not complex: the Not1-Caf1-Ccr4 interaction. Mol Cell 48(2):207–218
Taschner M, Bhogaraju S, Vetter M, Morawetz M, Lorentzen E (2011) Biochemical mapping of interactions within the Intraflagellar Transport (IFT) B core complex: IFT52 binds directly to four other IFT-B subunits. J Biol Chem 286:26344–26352
Lorentzen E et al (2005) The archaeal exosome core is a hexameric ring structure with three catalytic subunits. Nat Struct Mol Biol 12:575–581
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Basquin, J., Taschner, M., Lorentzen, E. (2016). Complex Reconstitution from Individual Protein Modules. In: Vega, M. (eds) Advanced Technologies for Protein Complex Production and Characterization. Advances in Experimental Medicine and Biology, vol 896. Springer, Cham. https://doi.org/10.1007/978-3-319-27216-0_19
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DOI: https://doi.org/10.1007/978-3-319-27216-0_19
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