title: One-step purification of the beta-glucan elicitor-binding protein from soybean (Glycine max L.) roots and characterization of an anti-peptide antiserum.
abstract: A low abundance beta-glucan elicitor-binding protein from soybean was isolated by a rapid, simple and one-step purification method yielding about 9000-fold enrichment. The affinity-based purification technique was more efficient than a procedure that uses conventional methods and preserved the binding activity to a much larger extent. The final preparation consisted of one major protein with an apparent molecular mass of about 75 kDa. Electrophoretic analyses of the purified and photoaffinity-labeled binding protein showed that the native protein was an oligomer with apparent molecular mass of about 240 kDa. A polyclonal anti-peptide antiserum was raised against a synthetic 15-mer internal oligopeptide sequence derived from the 75-kDa protein. The antiserum recognized the purified binding protein in immunoblotting experiments and precipitated the affinity-labeled protein from a crude extract of the membrane fraction.
authors.affiliation: Botanisches Institut der Universität, München, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Mithöfer
authors.forename: F
authors.initials: F
authors.lastname: Lottspeich
authors.forename: J
authors.initials: J
authors.lastname: Ebel
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D002846:Chromatography, Affinity / Q000379:methods; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005936:Glucans / Q000378:metabolism; D007106:Immune Sera; D007074:Immunoglobulin G; D037102:Lectins; D008861:Microsomes / Q000378:metabolism; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D018517:Plant Roots; D011817:Rabbits / Q000276:immunology; D013025:Glycine max / Q000378:metabolism*
chemical_list: D002352:Carrier Proteins; D005936:Glucans; D007106:Immune Sera; D007074:Immunoglobulin G; D037102:Lectins; D010446:Peptide Fragments; C037937:glucan-binding proteins