title: Interactions of Flagellar Structural Subunits with the Membrane Export Machinery.
abstract: During assembly of the bacterial flagellum, structural subunits synthesized inside the cell must be exported across the cytoplasmic membrane before they can crystallize into the nascent flagellar structure. This export process is facilitated by a specialized Flagellar Type III Secretion System (fT3SS) located at the base of each flagellum. Here, we describe three methods-isothermal titration calorimetry, photo-crosslinking using unnatural amino acids, and a subunit capture assay-used to investigate the interactions of flagellar structural subunits with the membrane export machinery component FlhB.
authors.affiliation: Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QP, UK.
authors.forename: Lewis D B
authors.initials: LD
authors.lastname: Evans
authors.affiliation: Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QP, UK.
authors.forename: Paul M
authors.initials: PM
authors.lastname: Bergen
authors.affiliation: Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QP, UK.
authors.forename: Owain J
authors.initials: OJ
authors.lastname: Bryant
authors.affiliation: Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QP, UK. gmf25@cam.ac.uk.
authors.forename: Gillian M
authors.initials: GM
authors.lastname: Fraser
mesh_terms: D001419:Bacteria / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism*; D005407:Flagella / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D021381:Protein Transport / Q000502:physiology*
chemical_list: D001426:Bacterial Proteins; D008565:Membrane Proteins
keywords: Capture assay; Flagella; Flagellar type III secretion system (fT3SS); Isothermal titration calorimetry (ITC); Protein export; Protein-protein interactions; Salmonella; p-benzoyl-L-phenylalanine (pBpa) photo-crosslinking