title: Deletion of the linker connecting the catalytic and cellulose-binding domains of endoglucanase A (CenA) of Cellulomonas fimi alters its conformation and catalytic activity.
abstract: The Pro-Thr box is a linker of 23 amino acids ((PT)4T(PT)7) connecting the catalytic domain and the cellulose-binding domain (CBD) of endoglucanase A (CenA) from the bacterium Cellulomonas fimi. Deletion of the Pro-Thr box alters the conformation of CenA by changing the relative orientation of the catalytic domain and the CBD. The tertiary structures of the catalytic domain and the CBD appear to be unchanged. The change in conformation reduces the catalytic efficiency of the enzyme and masks one of two protease-sensitive sites between the domains. The deletion does not affect the adsorption of the enzyme to microcrystalline cellulose, but it does affect its desorption from cellulose. The results suggest that the Pro-Thr box in CenA has an extended, kinked, and rigid conformation.
authors.affiliation: Department of Microbiology, University of British Columbia, Vancouver, Canada.
authors.forename: H
authors.initials: H
authors.lastname: Shen
authors.forename: M
authors.initials: M
authors.lastname: Schmuck
authors.forename: I
authors.initials: I
authors.lastname: Pilz
authors.forename: N R
authors.initials: NR
authors.lastname: Gilkes
authors.forename: D G
authors.initials: DG
authors.lastname: Kilburn
authors.forename: R C
authors.initials: RC
authors.lastname: Miller
authors.forename: R A
authors.initials: RA
authors.lastname: Warren
mesh_terms: D000192:Actinomycetales / Q000201:enzymology* / Q000235:genetics; D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D002480:Cellulase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D002482:Cellulose / Q000378:metabolism; D002872:Chromosome Deletion; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010957:Plasmids; D011487:Protein Conformation
chemical_list: D004269:DNA, Bacterial; D002482:Cellulose; D002480:Cellulase