title: Design, expression, and crystallization of recombinant lectin from the garden pea (Pisum sativum).
abstract: The propeptide form of the lectin from the garden pea (Pisum sativum agglutinin) has been expressed in Escherichia coli by attaching its cDNA to an inducible promoter. By a number of criteria, including the ability to form dimers, hemagglutination titer, Western blot, and enzyme-linked immunosorbent assay, the resulting propeptide molecule is virtually indistinguishable from the mature proteolytically processed lectin isolated from peas. Preliminary crystallization experiments using the recombinant propeptide lectin yield crystals in space group P2(1)2(1)2(1) with a = 64.8 A, b = 73.8 A, and c = 109.0 A (1 A = 0.1 nm) that diffract to 2.8-A resolution. This unit cell size is quite similar to the unit cell determined for native pea lectin, suggesting that the overall structure of the recombinant prolectin is virtually identical.
authors.affiliation: Department of Biochemistry, School of Medicine, University of Alabama, Birmingham 35294.
authors.forename: T
authors.initials: T
authors.lastname: Prasthofer
authors.forename: S R
authors.initials: SR
authors.lastname: Phillips
authors.forename: F L
authors.initials: FL
authors.lastname: Suddath
authors.forename: J A
authors.initials: JA
authors.lastname: Engler
mesh_terms: D000595:Amino Acid Sequence; D015153:Blotting, Western; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D003460:Crystallization; D004797:Enzyme-Linked Immunosorbent Assay; D006388:Hemagglutinins; D037102:Lectins / Q000235:genetics*; D008969:Molecular Sequence Data; D037121:Plant Lectins; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000648:ultrastructure
chemical_list: D006388:Hemagglutinins; D037102:Lectins; D037121:Plant Lectins; D011994:Recombinant Proteins; C036292:pea lectin