title: The crystal structure of Thermotoga maritima maltosyltransferase and its implications for the molecular basis of the novel transfer specificity.
abstract: Maltosyltransferase (MTase) from the hyperthermophile Thermotoga maritima represents a novel maltodextrin glycosyltransferase acting on starch and malto-oligosaccharides. It catalyzes the transfer of maltosyl units from alpha-1,4-linked glucans or malto-oligosaccharides to other alpha-1,4-linked glucans, malto-oligosaccharides or glucose. It belongs to the glycoside hydrolase family 13, which represents a large group of (beta/alpha)(8) barrel proteins sharing a similar active site structure. The crystal structures of MTase and its complex with maltose have been determined at 2.4 A and 2.1 A resolution, respectively. MTase is a homodimer, each subunit of which consists of four domains, two of which are structurally homologous to those of other family 13 enzymes. The catalytic core domain has the (beta/alpha)(8) barrel fold with the active-site cleft formed at the C-terminal end of the barrel. Substrate binding experiments have led to the location of two distinct maltose-binding sites; one lies in the active-site cleft, covering subsites -2 and -1; the other is located in a pocket adjacent to the active-site cleft. The structure of MTase, together with the conservation of active-site residues among family 13 glycoside hydrolases, are consistent with a common double-displacement catalytic mechanism for this enzyme. Analysis of maltose binding in the active site reveals that the transfer of dextrinyl residues longer than a maltosyl unit is prevented by termination of the active-site cleft after the -2 subsite by the side-chain of Lys151 and the stretch of residues 314-317, providing an explanation for the strict transfer specificity of MTase.
authors.affiliation: Krebs Institute for Biomolecular Research, Department of Molecular Biology and Biotechnology, The University of Sheffield, S10 2TN, England.
authors.forename: A
authors.initials: A
authors.lastname: Roujeinikova
authors.forename: C
authors.initials: C
authors.lastname: Raasch
authors.forename: J
authors.initials: J
authors.lastname: Burke
authors.forename: P J
authors.initials: PJ
authors.lastname: Baker
authors.forename: W
authors.initials: W
authors.lastname: Liebl
authors.forename: D W
authors.initials: DW
authors.lastname: Rice
mesh_terms: D001665:Binding Sites; D001692:Biological Transport; D018360:Crystallography, X-Ray; D019281:Dimerization; D005936:Glucans / Q000737:chemistry / Q000378:metabolism; D005964:Glucosyltransferases / Q000737:chemistry* / Q000378:metabolism*; D006004:Glycogen Debranching Enzyme System / Q000737:chemistry* / Q000378:metabolism*; D008320:Maltose / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D011487:Protein Conformation; D017510:Protein Folding; D013379:Substrate Specificity; D020124:Thermotoga maritima / Q000201:enzymology*
chemical_list: D005936:Glucans; D006004:Glycogen Debranching Enzyme System; D008320:Maltose; D005964:Glucosyltransferases; C436710:maltosyltransferase; C020569:4 alpha-glucanotransferase