Cytosolic proteins do not occur as isolated but are exposed to many interactions within a crowded cellular environment. We investigated the associations between a test cytosolic protein, human ileal bile acid binding protein (IBABP), and model cosolutes mimicking macromolecular and lipid membrane intracellular components. Using fluorescence spectroscopy, heteronuclear NMR, and molecular dynamics, we found that IBABP associated weakly with anionic lipid vesicles and experienced transient unspecific contacts with albumin. Localized dynamic perturbations were observed even in the case of apparent unspecific binding. IBABP and ubiquitin did not display mutually attractive forces, whereas IBABP associated specifically with lysozyme. A structural model of the IBABP-lysozyme complex was obtained by data-driven docking simulation. Presumably, all the interactions shown here contribute to modulating functional communication of a protein in its native environment.

Transient Interactions of a Cytosolic Protein with Macromolecular and Vesicular Cosolutes: Unspecific and Specific Effects

CECCON, Alberto;BUSATO, MIRKO;Perez Santero, Silvia;D'ONOFRIO, Mariapina;GIORGETTI, ALEJANDRO;ASSFALG, Michael
2015-01-01

Abstract

Cytosolic proteins do not occur as isolated but are exposed to many interactions within a crowded cellular environment. We investigated the associations between a test cytosolic protein, human ileal bile acid binding protein (IBABP), and model cosolutes mimicking macromolecular and lipid membrane intracellular components. Using fluorescence spectroscopy, heteronuclear NMR, and molecular dynamics, we found that IBABP associated weakly with anionic lipid vesicles and experienced transient unspecific contacts with albumin. Localized dynamic perturbations were observed even in the case of apparent unspecific binding. IBABP and ubiquitin did not display mutually attractive forces, whereas IBABP associated specifically with lysozyme. A structural model of the IBABP-lysozyme complex was obtained by data-driven docking simulation. Presumably, all the interactions shown here contribute to modulating functional communication of a protein in its native environment.
2015
NMR spectroscopy; data-driven docking; intracellular environment; macromolecular crowding; protein-protein interactions; Binding Sites; Cytosol; Humans; Hydroxysteroid Dehydrogenases; Lipid Bilayers; Macromolecular Substances; Molecular Dynamics Simulation; Muramidase; Nuclear Magnetic Resonance, Biomolecular; Protein Interaction Domains and Motifs; Spectrometry, Fluorescence; Ubiquitin
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11562/950222
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