Photocrosslinking approaches to interactome mapping

• Published in: Current opinion in chemical biology 2013-02, Vol.17 (1), p.90-101
• Main Authors: Pham, Nam D, Parker, Randy B, Kohler, Jennifer J
• Format: Article
• Language: English
• Subjects: Active Transport, Cell Nucleus; Animals; Cross-Linking Reagents - chemistry; Cross-Linking Reagents - metabolism; Humans; Photochemical Processes; Protein Binding; Protein Folding; Protein Interaction Mapping - methods; Proteins - chemistry; Proteins - genetics; Proteins - metabolism; Proteomics - methods; Signal Transduction; Transcriptional Activation
• ISSN: 1367-5931; 1879-0402
• DOI: 10.1016/j.cbpa.2012.10.034

[Display omitted] ► Photocrosslinking provides insight into context-dependent interactions. ► Interactome dynamics can be determined by photocrosslinking. ► Use of photocrosslinkers defines specific interaction interfaces within large complexes. ► Crosslinking captures transient interaction complexes containing IDPs and glycoproteins. 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.