Metals are employed by all types of organisms to perform a remarkable array of functions that are critical for life. The concentrations of many are highly regulated through homeostatic mechanisms and pathways through which organisms selectively and correctly metallate metalloproteins. Along these pathways, metal-mediated protein-protein interactions, i.e. those only occurring in the presence of the metal ion and through a direct interaction with it, contribute to select the correct route for metal transfer to the correct final destination; the metal is necessary for the interaction between the donor and the acceptor and, to prevent unproductive binding between the apo forms, no interaction occurs in its absence. The energetic contribution leading to the formation of detectable amounts of complex in the presence of the metal results from the involvement of amino acidic ligands from both protein partners in the coordination sphere of the metal ion. In the case of copper(I) trafficking pathways, copper-mediated protein-protein interactions have been conserved along evolution and can be exploited for different cellular functions, from copper incorporation into cellular organelle or copper enzymes in eukaryotes and bacteria, to metal detoxification in bacteria and regulatory role in the protein trafficking of mammal ATPases. NMR turns out to be the preferential tool to characterize these kinds of metal-mediated interactions, in such a way as being able to elucidate at the molecular level the above mentioned functional processes. The aim of this chapter is to present and discuss metal-mediated interactions, to address a NMR-based strategy for the structural determination of such complexes, which are usually in equilibrium with the free proteins with which they rapidly interconvert, and to provide protocols and troubleshooting. These aspects are illustrated with recent examples from our own research in studying copper-mediated protein-protein complexes.

Metal-Mediated Interactions / Simone, Ciofi-Baffoni. - STAMPA. - (2012), pp. 197-203. [10.1002/9783527644506.ch10]

Metal-Mediated Interactions

CIOFI BAFFONI, SIMONE
2012

Abstract

Metals are employed by all types of organisms to perform a remarkable array of functions that are critical for life. The concentrations of many are highly regulated through homeostatic mechanisms and pathways through which organisms selectively and correctly metallate metalloproteins. Along these pathways, metal-mediated protein-protein interactions, i.e. those only occurring in the presence of the metal ion and through a direct interaction with it, contribute to select the correct route for metal transfer to the correct final destination; the metal is necessary for the interaction between the donor and the acceptor and, to prevent unproductive binding between the apo forms, no interaction occurs in its absence. The energetic contribution leading to the formation of detectable amounts of complex in the presence of the metal results from the involvement of amino acidic ligands from both protein partners in the coordination sphere of the metal ion. In the case of copper(I) trafficking pathways, copper-mediated protein-protein interactions have been conserved along evolution and can be exploited for different cellular functions, from copper incorporation into cellular organelle or copper enzymes in eukaryotes and bacteria, to metal detoxification in bacteria and regulatory role in the protein trafficking of mammal ATPases. NMR turns out to be the preferential tool to characterize these kinds of metal-mediated interactions, in such a way as being able to elucidate at the molecular level the above mentioned functional processes. The aim of this chapter is to present and discuss metal-mediated interactions, to address a NMR-based strategy for the structural determination of such complexes, which are usually in equilibrium with the free proteins with which they rapidly interconvert, and to provide protocols and troubleshooting. These aspects are illustrated with recent examples from our own research in studying copper-mediated protein-protein complexes.
2012
9783527328505
NMR of Biomolecules. Towards Mechanistic Systems Biology
197
203
Simone, Ciofi-Baffoni
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Utilizza questo identificatore per citare o creare un link a questa risorsa: https://hdl.handle.net/2158/781463
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