title: Cloning and characterization of the Escherichia coli Heptosyltransferase III: Exploring substrate specificity in lipopolysaccharide core biosynthesis.
abstract: Bacterial lipopolysaccharide (LPS) molecules are an important cell surface component that enables adhesion to surfaces and cell motility, amongst other functions. In Escherichia coli, there are multiple Heptosyltransferase enzymes involved in the biosynthesis of the core region of LPS. Here we describe the first ever cloning, expression, purification and characterization of Heptosyltransferase III (HepIII) from E. coli, which catalyzes the addition of an L-glycero-D-manno-heptose (Hep) residue to the growing LPS core via an α(1→7) bond. Inspired by results from our lab on the E. coli HepI, we assessed the catalytic efficiency with phospho-Hep2-Kdo2-Lipid A (PH2K2LA) and two deacylated analogues.
authors.affiliation: Department of Chemistry, Wesleyan University, Middletown, CT 06459, USA.
authors.forename: Jagadesh
authors.initials: J
authors.lastname: Mudapaka
authors.affiliation: Department of Chemistry, Wesleyan University, Middletown, CT 06459, USA. Electronic address: eataylor@wesleyan.edu.
authors.forename: Erika Anne
authors.initials: EA
authors.lastname: Taylor
mesh_terms: D000595:Amino Acid Sequence; D055162:Biocatalysis; D002240:Carbohydrate Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016695:Glycosyltransferases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006539:Heptoses / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D008070:Lipopolysaccharides / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D015394:Molecular Structure; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
chemical_list: D029968:Escherichia coli Proteins; D006539:Heptoses; C506188:Kdo2-lipid A; D008070:Lipopolysaccharides; C026312:glycero-manno-heptose; D016695:Glycosyltransferases; C409702:WaaQ protein, E coli
keywords: Bacterial biofilms; Core oligosaccharide; Glycosyltransferase; Heptosyltransferase; Lipopolysaccharide biosynthesis