title: Isolation and characterization of pro-barley lectin expressed in Escherichia coli.
abstract: Lectins are a class of proteins with specific carbohydrate-binding properties found in a wide variety of plants and animals. Gramineae lectins are presumably defense-related proteins in plants that exert their effect by binding to N-acetylglucosamine. Barley lectin is a vacuolar protein synthesized with an amino-terminal signal sequence for entering the secretory pathway and a carboxyl-terminal propeptide necessary for proper targeting to the vacuole. To analyze the three-dimensional structure of barley lectin with the carboxyl-terminal extension and to investigate whether the conversion of the prolectin into the mature molecule leads to a conformational change, the precursor and the mature forms of barley lectin were expressed in Escherichia coli. Both proteins accumulated in denatured form in inclusion bodies were solubilized in 8 M urea and renatured in a redox buffer system. Active pro- and mature barley lectins were purified to homogeneity by affinity chromatography.
authors.affiliation: Department of Energy Plant Research Laboratory, Michigan State University, East Lansing 48824-1312.
authors.forename: M R
authors.initials: MR
authors.lastname: Schroeder
authors.forename: N V
authors.initials: NV
authors.lastname: Raikhel
mesh_terms: D000117:Acetylglucosamine / Q000378:metabolism; D001483:Base Sequence; D015153:Blotting, Western; D002846:Chromatography, Affinity / Q000379:methods; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D001467:Hordeum / Q000737:chemistry* / Q000235:genetics; D002479:Inclusion Bodies; D037102:Lectins / Q000096:biosynthesis / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D037121:Plant Lectins; D011489:Protein Denaturation; D011498:Protein Precursors / Q000096:biosynthesis / Q000187:drug effects / Q000302:isolation & purification*; D014508:Urea / Q000494:pharmacology
chemical_list: D037102:Lectins; D037121:Plant Lectins; D011498:Protein Precursors; C076978:barley lectin; D014508:Urea; D000117:Acetylglucosamine