title: Synthesis of a stable analog of the phosphorylated form of CheY: phosphono-CheY.
abstract: The chemical modification of a cysteinyl residue of D57C CheY by the addition of a phosphonomethyl group, (HO)(2)P(O)-CH(2)-, is described. This modification produces a nonlabile analog of an aspartyl phosphate residue in the active form of CheY. The chemically modified protein, phosphono-CheY, is suitable for structural and functional studies. An extensive discussion of the synthetic methodology and purification strategy is presented. A detailed protocol is given.
authors.affiliation: Department of Chemistry and Biochemistry, University of North Carolina, Wilmington, North Carolina, USA.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Halkides
authors.forename: Cory J
authors.initials: CJ
authors.lastname: Bottone
authors.forename: Eric S
authors.initials: ES
authors.lastname: Casper
authors.forename: R Matthew
authors.initials: RM
authors.lastname: Haas
authors.forename: Kenneth
authors.initials: K
authors.lastname: McAdams
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D019921:Biotinylation; D002851:Chromatography, High Pressure Liquid; D004228:Dithionitrobenzoic Acid; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D000072236:Methyl-Accepting Chemotaxis Proteins; D063065:Organophosphonates / Q000737:chemistry*; D010750:Phosphoproteins / Q000737:chemistry; D010766:Phosphorylation
chemical_list: D001426:Bacterial Proteins; D008565:Membrane Proteins; D000072236:Methyl-Accepting Chemotaxis Proteins; D063065:Organophosphonates; D010750:Phosphoproteins; C005149:phosphonoproteins; D004228:Dithionitrobenzoic Acid