title: Purification of a protein from Agrobacterium tumefaciens strain A348 that binds phenolic compounds.
abstract: In order to induce tumours on dicotyledonous plants, the bacterium Agrobacterium tumefaciens needs to be able to sense signal molecules, i.e. phenolic compounds. In order to identify putative chemoreceptors or environmental sensors involved in vir gene induction, we undertook the purification of a phenol-binding protein by affinity chromatography on a syringamide Ultrogel A4 column equilibrated at pH 5.6. A mild extraction of bacterial proteins with a Tris/HCl buffer at pH 9.0 led to the purification of a 39 kDa protein (Pbp39) with a pl of 4.3 after specific elution of the affinity matrix with sodium syringate. When the affinity chromatography was performed at neutral pH, barely any protein was isolated, indicating the importance of an acidic pH for optimal affinity. A microplate binding experiment revealed that both syringlyl biotinylated-BSA and sinapyl-biotinylated-BSA bound at pH 5.6 to the plate coated with Pbp39.
authors.affiliation: Centre de Biophysique Moléculaire, C.N.R.S, Orléans, France.
authors.forename: F
authors.initials: F
authors.lastname: Dyé
authors.forename: F M
authors.initials: FM
authors.lastname: Delmotte
mesh_terms: D016960:Agrobacterium tumefaciens / Q000737:chemistry* / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D005707:Gallic Acid / Q000031:analogs & derivatives; D008970:Molecular Weight; D010636:Phenols / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding
chemical_list: D001426:Bacterial Proteins; D010636:Phenols; D005707:Gallic Acid; C001945:syringic acid