title: The structural bases of the processive degradation of iota-carrageenan, a main cell wall polysaccharide of red algae.
abstract: iota-Carrageenans are sulfated 1,3-alpha-1,4-beta-galactans from the cell walls of red algae, which auto-associate into crystalline fibers made of aggregates of double-stranded helices. iota-Carrageenases, which constitute family 82 of glycoside hydrolases, fold into a right-handed beta-helix. Here, the structure of Alteromonas fortis iota-carrageenase bound to iota-carrageenan fragments was solved at 2.0A resolution (PDB 1KTW). The enzyme holds a iota-carrageenan tetrasaccharide (subsites +1 to +4) and a disaccharide (subsites -3, -4), thus providing the first direct determination of a 3D structure of iota-carrageenan. Electrostatic interactions between basic protein residues and the sulfate substituents of the polysaccharide chain dominate iota-carrageenan recognition. Glu245 and Asp247 are the proton donor and the base catalyst, respectively. C-terminal domain A, which was highly flexible in the native enzyme structure, adopts a alpha/beta-fold, also found in DNA/RNA-binding domains. In the substrate-enzyme complex, this polyanion-binding module shifts toward the beta-helix groove, forming a tunnel. Thus, from an open conformation which allows for the initial endo-attack of iota-carrageenan chains, the enzyme switches to a closed-tunnel form, consistent with its highly processive character, as seen from the electron-microscopy analysis of the degradation of iota-carrageenan fibers.
authors: Végétaux Marins et Biomolécules, UMR 7139 (CNRS/UPMC/Laboratories Goëmar), Station Biologique de Roscoff, Place Georges Teissier, BP 74, 29682 Roscoff Cedex, Brittany, France.kloareg@sb-rosecoff.ff
authors: Gurvan
authors: G
authors: Michel
authors: William
authors: W
authors: Helbert
authors: Richard
authors: R
authors: Kahn
authors: Otto
authors: O
authors: Dideberg
authors: Bernard
authors: B
authors: Kloareg
mesh_terms: D020591:Alteromonas / Q000201:enzymology; D001426:Bacterial Proteins / Q000378:metabolism*; D001665:Binding Sites; D002351:Carrageenan / Q000737:chemistry* / Q000378:metabolism*; D020134:Catalytic Domain; D002473:Cell Wall / Q000737:chemistry / Q000378:metabolism*; D003461:Crystallography; D006026:Glycoside Hydrolases / Q000378:metabolism*; D008958:Models, Molecular; D011485:Protein Binding; D017510:Protein Folding; D017433:Protein Structure, Secondary; D000461:Rhodophyta / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D002351:Carrageenan; D006026:Glycoside Hydrolases; C019846:kappa-carrageenase protein, Alteromonadaceae