The cellulose‐binding domain of endoglucanase A (CenA) from <i>Cellulomonas fimi</i>: evidence for the involvement of tryptophan residues in binding

Din, Neena; Forsythe, Ian J.; Burtnick, Leslie D.; Gilkes, Neil R.; Miller, Robert C.; Warren, R. A. J.; Kilburn, Douglas G.

doi:10.1111/j.1365-2958.1994.tb00352.x

Cited by 133 publications

(111 citation statements)

References 43 publications

(17 reference statements)

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“…Consistent with earlier investigations (7,12,25,26), our current data confirm that tryptophan residues contribute higher energy of binding than tyrosines on a CBM binding surface. However, the effect is limited to one Trp residue; addition of two Trp residues led to no further improvement in binding.…”

Section: Discussionsupporting

confidence: 82%

The binding specificity and affinity determinants of family 1 and family 3 cellulose binding modules

Lehtiö

Sugiyama

Gustavsson

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

312

259

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Cellulose binding modules (CBMs) potentiate the action of cellulolytic enzymes on insoluble substrates. Numerous studies have established that three aromatic residues on a CBM surface are needed for binding onto cellulose crystals and that tryptophans contribute to higher binding affinity than tyrosines. However, studies addressing the nature of CBM-cellulose interactions have so far failed to establish the binding site on cellulose crystals targeted by CBMs. In this study, the binding sites of CBMs on Valonia cellulose crystals have been visualized by transmission electron microscopy. Fusion of the CBMs with a modified staphylococcal protein A (ZZ-domain) allowed direct immuno-gold labeling at close proximity of the actual CBM binding site. The transmission electron microscopy images provide unequivocal evidence that the fungal family 1 CBMs as well as the family 3 CBM from Clostridium thermocellum CipA have defined binding sites on two opposite corners of Valonia cellulose crystals. In most samples these corners are worn to display significant area of the hydrophobic (110) plane, which thus constitutes the binding site for these CBMs.

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Section: Discussionsupporting

confidence: 82%

The binding specificity and affinity determinants of family 1 and family 3 cellulose binding modules

Lehtiö

Sugiyama

Gustavsson

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

312

259

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“…cellulosa (Poole et al, 1993), Ala substitutions of the Trp residues resulted in a considerable decrease in the binding capacity to cellulose, whereas mutant proteins containing Phe substitutions retained some of their binding ability. However, the exchange of either of the conserved Trp residues in the CBD of the ~~l~u~~~ endoglucanase (CenA) led to differently pronounced effects on the binding capacities of the mutated proteins (Din et al, 1994).…”

Section: Discussionmentioning

confidence: 99%

Specific Interaction of the Streptomyces Chitin‐Binding Protein Chb1 with α‐Chitin

Zeltiņš

Schrempp

1997

European Journal of Biochemistry

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Streptomyces olivaceoviridis secretes a so far unique protein of 18.7 kDa (CHB1) which lacks catalytic activity. It interacts highly specifically with a-chitin, but not with @-chitin, chitosan, or cellulose. Each of the five codons for tryptophan (Trp) in the chbl gene was replaced by those for leucine (Leu) or tyrosine (Tyr). Eight corresponding mutant proteins and the wild-type protein were purified to homogeneity and their binding capacity to a-chitin was determined. The relative affinities to anti-CHB1 antibodies, the kinetics of binding, the dissociation constants, circular dichroism, and fluorescence emission spectra for three mutant types were compared to the characteristics of CHB1. The presented data lead to the following conclusions.

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“…(61). Mutation of tryptophan residues to alanyl residues in CBDCenA significantly decreases the affinity of the polypeptide for cellulose without perturbation of protein folding (9). The other explanation may be that SngPT-CBDc,x (29).…”

Section: Methodsmentioning

confidence: 99%

Streptomyces lividans glycosylates the linker region of a beta-1,4-glycanase from Cellulomonas fimi

et al. 1994

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The 0-1,4-glycanase Cex of the gram-positive bacterium Cellulomonas fimi is a glycoprotein comprising a C-terminal cellulose-binding domain connected to an N-terminal catalytic domain by a linker containing only prolyl and threonyl (PT) residues. Cex is also glycosylated by Streptomyces lividans. The glycosylation of Cex produced in both C. fimi and S. lividans protects the enzyme from proteolysis. When the gene fragments encoding the cellulose-binding domain of Cex (CBDCex), the PT linker plus CBDCex (PT-CBDcex), and the catalytic domain plus CBDCeX of Cex were expressed in S. lividans, only PT-CBDCex was glycosylated. Therefore, all the glycans must be 0 linked because only the PT linker was glycosylated. A glycosylated form and a nonglycosylated form of PT-CBDCex were produced by S. lividans. The glycosylated form of PT-CBDCex was heterogeneous; its average carbohydrate content was -10 mol of D-mannose equivalents per mol of protein, but the glycans contained from 4 to 12 a-D-mannosyl and a-D-galactosyl residues. Glycosylated Cex from S. lividans was also heterogeneous. The presence of glycans on PT-CBDCex increased its affinity for bacterial microcrystalline cellulose. The location of glycosylation only on the linker region of Cex correlates with the properties conferred on the enzyme by the glycans.Cellulomonas fimi is a gram-positive coryneform bacterium.It produces several cellulases, two of which, endo-,B-1,4-glucanase A (CenA) and a J-1,4-xylanase/exo-P3-1,4-glucanase

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The cellulose‐binding domain of endoglucanase A (CenA) from Cellulomonas fimi: evidence for the involvement of tryptophan residues in binding

Cited by 133 publications

References 43 publications

The binding specificity and affinity determinants of family 1 and family 3 cellulose binding modules

The binding specificity and affinity determinants of family 1 and family 3 cellulose binding modules

Specific Interaction of the Streptomyces Chitin‐Binding Protein Chb1 with α‐Chitin

Streptomyces lividans glycosylates the linker region of a beta-1,4-glycanase from Cellulomonas fimi

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