Carbohydrate Binding Module Family 28

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• Author: ^^^Shinya Fushinobu^^^
• Responsible Curator: ^^^Shinya Fushinobu^^^

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CAZy DB link

http://www.cazy.org/CBM28.html

Ligand specificities

This module was firstly classified in 2002 by a study of the C-terminal module in CH5 Cel5A from Bacillus sp. 1139 (BspCBM28) [1]. Currently known CBM28s are solely from bacterial origins, mostly attached to cellulosomal endoglucanases in tandem with CBM17. They bind non-crystalline (or amorphous) part of cellulose, cellooligosaccharides, or β-1,3-1,4-glucans [2].

Structural Features

Figure 1: CBM28 (3aci) [3] in comparison with CBM17 (1j84) [4] and CBM4 (1gu3) [5]. Sugars of cellooligosaccharides (cellopentaose or cellotetraose) are designated as A-D (+E) from the non-reducing end to the reducing end.

• Fold: CBM28s have a β-sandwich fold (approximately 200 amino acids) that is similar to CBM17 and CBM4 (Figure 1). The module has a straight cleft that binds a cellulose glycan chain at the center of the β-sandwich fold.
• Type: CBM28s are typical endo-type Type C CBMs that accommodate a single glycan chain because both ends of the cleft are open.
• Features of ligand binding: Figure 1 shows the cellopentaose complex structure (3aci) of CBM28 in GH5 Cel5A from Clostridium josui (CjCBM28). The long binding cleft runs at the center of the molecule. Therefore, the binding pocket location of CBM28 is on a face (side) of the β-sandwich fold (CBM Fold). The shallow cleft of CBM28 binds one side of the cellooligosaccharides (face-on) in contrast with the case of CBM4 (side-on). There are five subsites (A-E from the non-reducing end to the reducing end) in CjCBM28. Interestingly, the direction of the cellooligosaccharides bound to CBM28 is opposite to those in CBM17 and CBM4. Subsites B, C, and E form stacking interactions with aromatic residues (W78, W129, and F128 in CjCBM28). The flanking hydroxyl groups are extensively recognized by direct or water-mediated hydrogen bonds. Therefore, CBM28 has a relatively wide cleft.

Functionalities

• Functional role of CBM: CBM28s are thought to target catalytic modules (endoglucanases) to non-crystalline region of cellulose. Deletion mutants of CBM28 in Cel5A from Bacillus sp. 1139 significantly decreased amounts of the soluble products from amorphous cellulose [2].

Describe common functional roles such as targeting, disruptive, anchoring, proximity/position on substrate.

• Most Common Associated Modules: Endo-β-1,4-glucanases of GH5. A survey using the GH-CBM tool shows that CBM28s are not associated with other GH family domains.
• Novel Applications: BspCBM28 was used as a most amorphous-specific CBM in a study of photo activated localization microscopy (PALM) [6].

Family Firsts

First Identified

The first identified member is the C-terminal module in Cel5A from Bacillus sp. 1139 (BspCBM28) [1]

First Structural Characterization

Partial assignment of the NMR data of a CBM28 in Cel5I from Clostridium cellulolyticum was reported in 2002 [7] but its three-dimensional structure is not reported yet. The first crystal structure was reported in 2004 for BspCBM28 in a ligand-free form 1uww [8]. The first ligand complex structures were reported in 2010 for CBM28 in Cel5A from C. josui (CjCBM28) 3acf 3acg 3ach 3aci.

References

1. Boraston AB, Ghaffari M, Warren RA, and Kilburn DG. (2002). Identification and glucan-binding properties of a new carbohydrate-binding module family. Biochem J. 2002;361(Pt 1):35-40. DOI:10.1042/0264-6021:3610035 |
2. Boraston AB, Kwan E, Chiu P, Warren RA, and Kilburn DG. (2003). Recognition and hydrolysis of noncrystalline cellulose. J Biol Chem. 2003;278(8):6120-7. DOI:10.1074/jbc.M209554200 |
3. Tsukimoto K, Takada R, Araki Y, Suzuki K, Karita S, Wakagi T, Shoun H, Watanabe T, and Fushinobu S. (2010). Recognition of cellooligosaccharides by a family 28 carbohydrate-binding module. FEBS Lett. 2010;584(6):1205-11. DOI:10.1016/j.febslet.2010.02.027 |
4. Notenboom V, Boraston AB, Chiu P, Freelove AC, Kilburn DG, and Rose DR. (2001). Recognition of cello-oligosaccharides by a family 17 carbohydrate-binding module: an X-ray crystallographic, thermodynamic and mutagenic study. J Mol Biol. 2001;314(4):797-806. DOI:10.1006/jmbi.2001.5153 |
5. Boraston AB, Nurizzo D, Notenboom V, Ducros V, Rose DR, Kilburn DG, and Davies GJ. (2002). Differential oligosaccharide recognition by evolutionarily-related beta-1,4 and beta-1,3 glucan-binding modules. J Mol Biol. 2002;319(5):1143-56. DOI:10.1016/S0022-2836(02)00374-1 |
6. Fox JM, Jess P, Jambusaria RB, Moo GM, Liphardt J, Clark DS, and Blanch HW. (2013). A single-molecule analysis reveals morphological targets for cellulase synergy. Nat Chem Biol. 2013;9(6):356-61. DOI:10.1038/nchembio.1227 |
7. Mosbah A, Tardif C, Bornet O, Valette O, Henrissat B, and Darbon H. (2002). Assignment of the 1H, 13C, and 15N resonances of the 22,5 kDa CBM28 module of the cellulase Cel5I of Clostridium cellulolyticum. J Biomol NMR. 2002;23(2):157-8. DOI:10.1023/a:1016354623398 |
8. Jamal S, Nurizzo D, Boraston AB, and Davies GJ. (2004). X-ray crystal structure of a non-crystalline cellulose-specific carbohydrate-binding module: CBM28. J Mol Biol. 2004;339(2):253-8. DOI:10.1016/j.jmb.2004.03.069 |