Importance of Hydrophobic and Polar Residues in Ligand Binding in the Family 15 Carbohydrate-Binding Module fromCellvibrio japonicusXyn10C

Biochemistry ◽  
2003 ◽  
Vol 42 (31) ◽  
pp. 9316-9323 ◽  
Author(s):  
Gavin Pell ◽  
Michael P. Williamson ◽  
Christopher Walters ◽  
Haomao Du ◽  
Harry J. Gilbert ◽  
...  
Keyword(s):  
Ligand Binding ◽  
Carbohydrate Binding ◽  
Carbohydrate Binding Module ◽  
The Family ◽  
Polar Residues

Crystal Structures of the Family 9 Carbohydrate-Binding Module fromThermotoga maritimaXylanase 10A in Native and Ligand-Bound Forms†,‡

Biochemistry ◽  
2001 ◽  
Vol 40 (21) ◽  
pp. 6248-6256 ◽  
Author(s):  
Valerie Notenboom ◽  
Alisdair B. Boraston ◽  
Douglas G. Kilburn ◽  
David R. Rose
Keyword(s):  
Crystal Structures ◽  
Carbohydrate Binding ◽  
Carbohydrate Binding Module ◽  
The Family

Clostridium thermocellumXyn10B Carbohydrate-Binding Module 22-2:  The Role of Conserved Amino Acids in Ligand Binding†,‡

Biochemistry ◽  
2001 ◽  
Vol 40 (31) ◽  
pp. 9167-9176 ◽  
Author(s):  
Hefang Xie ◽  
Harry J. Gilbert ◽  
Simon J. Charnock ◽  
Gideon J. Davies ◽  
Michael P. Williamson ◽  
...  
Keyword(s):  
Amino Acids ◽  
Ligand Binding ◽  
Carbohydrate Binding ◽  
Carbohydrate Binding Module ◽  
Conserved Amino Acids

scholarly journals Novel xylan-binding properties of an engineered family 4 carbohydrate-binding module

2007 ◽  
Vol 406 (2) ◽  
pp. 209-214 ◽  
Author(s):  
Lavinia Cicortas Gunnarsson ◽  
Cedric Montanier ◽  
Richard B. Tunnicliffe ◽  
Mike P. Williamson ◽  
Harry J. Gilbert ◽  
...  
Keyword(s):  
Ligand Binding ◽  
Binding Site ◽  
Hydrophobic Interactions ◽  
Molecular Engineering ◽  
Carbohydrate Binding ◽  
Carbohydrate Binding Module ◽  
Wild Type ◽  
Rhodothermus Marinus ◽  
Binding Properties ◽  
Carbohydrate Binding Modules

Molecular engineering of ligand-binding proteins is commonly used for identification of variants that display novel specificities. Using this approach to introduce novel specificities into CBMs (carbohydrate-binding modules) has not been extensively explored. Here, we report the engineering of a CBM, CBM4-2 from the Rhodothermus marinus xylanase Xyn10A, and the identification of the X-2 variant. As compared with the wild-type protein, this engineered module displays higher specificity for the polysaccharide xylan, and a lower preference for binding xylo-oligomers rather than binding the natural decorated polysaccharide. The mode of binding of X-2 differs from other xylan-specific CBMs in that it only has one aromatic residue in the binding site that can make hydrophobic interactions with the sugar rings of the ligand. The evolution of CBM4-2 has thus generated a xylan-binding module with different binding properties to those displayed by CBMs available in Nature.

scholarly journals Molecular basis for the preferential recognition of β1,3‐1,4‐glucans by the family 11 carbohydrate‐binding module from Clostridium thermocellum

FEBS Journal ◽  
2019 ◽  
Vol 287 (13) ◽  
pp. 2723-2743 ◽  
Author(s):  
Diana O. Ribeiro ◽  
Aldino Viegas ◽  
Virgínia M. R. Pires ◽  
João Medeiros‐Silva ◽  
Pedro Bule ◽  
...  
Keyword(s):  
Molecular Basis ◽  
Clostridium Thermocellum ◽  
Carbohydrate Binding ◽  
Carbohydrate Binding Module ◽  
The Family ◽  
Preferential Recognition

scholarly journals The Family 6 Carbohydrate Binding ModuleCmCBM6-2 Contains Two Ligand-binding Sites with Distinct Specificities

2004 ◽  
Vol 279 (20) ◽  
pp. 21552-21559 ◽  
Author(s):  
Joanna L. Henshaw ◽  
David N. Bolam ◽  
Virgínia M. R. Pires ◽  
Mirjam Czjzek ◽  
Bernard Henrissat ◽  
...  
Keyword(s):  
Ligand Binding ◽  
Binding Sites ◽  
Carbohydrate Binding ◽  
The Family ◽  
Ligand Binding Sites

scholarly journals The family 21 carbohydrate-binding module of glucoamylase from Rhizopus oryzae consists of two sites playing distinct roles in ligand binding

2006 ◽  
Vol 396 (3) ◽  
pp. 469-477 ◽  
Author(s):  
Wei-I Chou ◽  
Tun-Wen Pai ◽  
Shi-Hwei Liu ◽  
Bor-Kai Hsiung ◽  
Margaret D.-T. Chang
Keyword(s):  
Ligand Binding ◽  
Binding Sites ◽  
Catalytic Domain ◽  
Rhizopus Oryzae ◽  
Structural Information ◽  
Molecular Model ◽  
Site Directed Mutagenesis ◽  
Carbohydrate Binding ◽  
Carbohydrate Binding Module ◽  
Ligand Binding Sites

The starch-hydrolysing enzyme GA (glucoamylase) from Rhizopus oryzae is a commonly used glycoside hydrolase in industry. It consists of a C-terminal catalytic domain and an N-terminal starch-binding domain, which belong to the CBM21 (carbohydrate-binding module, family 21). In the present study, a molecular model of CBM21 from R. oryzae GA (RoGACBM21) was constructed according to PSSC (progressive secondary structure correlation), modified structure-based sequence alignment, and site-directed mutagenesis was used to identify and characterize potential ligand-binding sites. Our model suggests that RoGACBM21 contains two ligand-binding sites, with Tyr32 and Tyr67 grouped into site I, and Trp47, Tyr83 and Tyr93 grouped into site II. The involvement of these aromatic residues has been validated using chemical modification, UV difference spectroscopy studies, and both qualitative and quantitative binding assays on a series of RoGACBM21 mutants. Our results further reveal that binding sites I and II play distinct roles in ligand binding, the former not only is involved in binding insoluble starch, but also facilitates the binding of RoGACBM21 to long-chain soluble polysaccharides, whereas the latter serves as the major binding site mediating the binding of both soluble polysaccharide and insoluble ligands. In the present study we have for the first time demonstrated that the key ligand-binding residues of RoGACBM21 can be identified and characterized by a combination of novel bioinformatics methodologies in the absence of resolved three-dimensional structural information.

Sign in / Sign up

Export Citation Format

Share Document