title: Photocrosslinking approaches to interactome mapping.
abstract: Photocrosslinking approaches can be used to map interactome networks within the context of living cells. Photocrosslinking methods rely on use of metabolic engineering or genetic code expansion to incorporate photocrosslinking analogs of amino acids or sugars into cellular biomolecules. Immunological and mass spectrometry techniques are used to analyze crosslinked complexes, thereby defining specific interactomes. Because photocrosslinking can be conducted in native, cellular settings, it can be used to define context-dependent interactions. Photocrosslinking methods are also ideally suited for determining interactome dynamics, mapping interaction interfaces, and identifying transient interactions in which intrinsically disordered proteins and glycoproteins engage. Here we discuss the application of cell-based photocrosslinking to the study of specific problems in immune cell signaling, transcription, membrane protein dynamics, nucleocytoplasmic transport, and chaperone-assisted protein folding.
authors.affiliation: Department of Biochemistry, 5323 Harry Hines Boulevard, University of Texas Southwestern Medical Center, Dallas, TX 75390-9038, USA.
authors.forename: Nam D
authors.initials: ND
authors.lastname: Pham
authors.forename: Randy B
authors.initials: RB
authors.lastname: Parker
authors.forename: Jennifer J
authors.initials: JJ
authors.lastname: Kohler
mesh_terms: D021581:Active Transport, Cell Nucleus; D000818:Animals; D003432:Cross-Linking Reagents / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D055668:Photochemical Processes; D011485:Protein Binding; D017510:Protein Folding; D025941:Protein Interaction Mapping / Q000379:methods*; D011506:Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D015398:Signal Transduction; D015533:Transcriptional Activation
chemical_list: D003432:Cross-Linking Reagents; D011506:Proteins