In vivo crosslinking methods for analyzing the assembly and architecture of chemoreceptor arrays.
The chemoreceptor molecules that mediate chemotactic responses in bacteria and archaea are physically clustered and operate as highly cooperative arrays. Few experimental approaches are able to investigate the structure-function organization of these chemoreceptor networks in living cells. This chapter describes chemical crosslinking methods that can be applied under normal physiological conditions to explore physical interactions between chemoreceptors and their underlying genetic and structural basis. Most of these crosslinking approaches are based on available atomic structures for chemoreceptor homodimers, the fundamental building block for higher-order networks. However, the general logic of our in vivo crosslinking approaches is readily applicable to other protein-protein interactions and other organisms, even when high-resolution structural information is not available.
Studdert
Claudia A
CA
Instituto de Investigaciones Biológicas, Universidad Nacional de Mar del Plata, Mar del Plata, Argentina.
Parkinson
John S
JS
D019843:Archaeal Proteins / Q000737:chemistry; D001419:Bacteria / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry; D001671:Biochemistry / Q000379:methods*; D002628:Chemoreceptor Cells / Q000737:chemistry*; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D003545:Cysteine / Q000737:chemistry; D003593:Cytoplasm / Q000378:metabolism; D019281:Dimerization; D004220:Disulfides / Q000737:chemistry; D029968:Escherichia coli Proteins / Q000737:chemistry; D008239:Lysine / Q000737:chemistry; D008956:Models, Chemical; D008957:Models, Genetic; D008968:Molecular Conformation; D009154:Mutation; D011485:Protein Binding
D019843:Archaeal Proteins; D001426:Bacterial Proteins; D003432:Cross-Linking Reagents; D004220:Disulfides; D029968:Escherichia coli Proteins; D008239:Lysine; D003545:Cysteine