title: Interaction between Clostridium thermocellum endoglucanase CelD and polypeptides derived from the cellulosome-integrating protein CipA: stoichiometry and cellulolytic activity of the complexes.
abstract: Four mini-scaffoldins were constructed from modules derived from the Clostridium thermocellum cellulosome-integrating protein CipA. Cip7 and Cip6 contained one and two cohesin modules respectively. Cip14 and Cip16, also containing one and two cohesin modules respectively, were flanked by a cellulose-binding domain. Endoglucanase CelD formed stable complexes with all mini-scaffoldins. Analytical ultracentrifugation of the complexes showed that 1 mol of CelD bound per mol of Cip14, and 2 mol of CelD bound per mol of Cip16. Under the conditions used for assaying cellulase activity, 96% of CelD alone bound to Avicel. Association with Cip14 or Cip16 increased the cellulose binding of CelD to 99%, while association with Cip7 or Cip6 decreased binding to 79 and 75% respectively. The hydrolytic activity of CelD against Avicel was increased 3-fold in complexes with Cip14 and Cip16, but remained substantially the same in complexes with Cip6 and Cip7. Addition of whole CipA also enhanced the efficiency of Avicel hydrolysis by CelD. However, even at an optimal ratio of the components, CelD-CipA complexes were somewhat less active than complexes of CelD with Cip14 or Cip16. These results suggest that the synergism observed between CelD and Cip14 or Cip16 is mostly due to the presence of the cellulose-binding domain, which promotes productive binding of the enzyme.
authors.affiliation: Unité de Physiologie Cellulaire, URA 1300 CNRS, Institut Pasteur, Paris, France.
authors.forename: I
authors.initials: I
authors.lastname: Kataeva
authors.forename: G
authors.initials: G
authors.lastname: Guglielmi
authors.forename: P
authors.initials: P
authors.lastname: Béguin
mesh_terms: D000327:Adsorption; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D002480:Cellulase / Q000737:chemistry / Q000378:metabolism*; D002482:Cellulose / Q000378:metabolism; D003013:Clostridium / Q000737:chemistry / Q000201:enzymology*; D006868:Hydrolysis; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D009097:Multienzyme Complexes / Q000737:chemistry / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; C080369:CipA protein, Clostridium; D008565:Membrane Proteins; D009097:Multienzyme Complexes; D010455:Peptides; D002482:Cellulose; C087879:endoglucanase CelD; D002480:Cellulase