title: Visualizing protein partnerships in living cells and organisms.
abstract: In recent years, scientists have expanded their focus from cataloging genes to characterizing the multiple states of their translated products. One anticipated result is a dynamic map of the protein association networks and activities that occur within the cellular environment. While in vitro-derived network maps can illustrate which of a multitude of possible protein-protein associations could exist, they supply a falsely static picture lacking the subtleties of subcellular location (where) or cellular state (when). Generating protein association network maps that are informed by both subcellular location and cell state requires novel approaches that accurately characterize the state of protein associations in living cells and provide precise spatiotemporal resolution. In this review, we highlight recent advances in visualizing protein associations and networks under increasingly native conditions. These advances include second generation protein complementation assays (PCAs), chemical and photo-crosslinking techniques, and proximity-induced ligation approaches. The advances described focus on background reduction, signal optimization, rapid and reversible reporter assembly, decreased cytotoxicity, and minimal functional perturbation. Key breakthroughs have addressed many challenges and should expand the repertoire of tools useful for generating maps of protein interactions resolved in both time and space.
authors: Yale University, Department of Molecular Biophysics and Biochemistry, 60 Whitney Ave., New Haven, CT 06520-8114, USA.
authors: Melissa A
authors: MA
authors: Lowder
authors: Jacob S
authors: JS
authors: Appelbaum
authors: Elissa M
authors: EM
authors: Hobert
authors: Alanna
authors: A
authors: Schepartz
mesh_terms: D003432:Cross-Linking Reagents / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000138:chemical synthesis* / Q000378:metabolism; D008856:Microscopy, Fluorescence; D055668:Photochemical Processes / Q000528:radiation effects; D010777:Photochemistry / Q000379:methods*; D025941:Protein Interaction Mapping / Q000379:methods*; D060066:Protein Interaction Maps; D011506:Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D013194:Staining and Labeling / Q000379:methods*; D014466:Ultraviolet Rays
chemical_list: D003432:Cross-Linking Reagents; D005456:Fluorescent Dyes; D011506:Proteins