Site-specific characterization of the association of xylooligosaccharides with the CBM13 lectin-like xylan binding domain from Streptomyces lividans xylanase 10A by NMR spectroscopy

Biochemistry
Manuela SchärpfLawrence P McIntosh

Abstract

Endo-beta-1,4-xylanase 10A (Xyn10A) from Streptomyces lividans includes an N-terminal catalytic module and a 130-residue C-terminal family 13 carbohydrate-binding module (CBM13). This latter domain adopts a beta-trefoil structure with three potential binding sites (alpha, beta, and gamma) for a variety of small sugars, xylooligosaccharides, and xylan polymers. To investigate the role of this multivalency in carbohydrate binding, we have used NMR spectroscopy to characterize the interaction of isolated CBM13 with a series of sugars. We have assigned resonances from the main chain nuclei of CBM13 using heteronuclear NMR experiments. Analysis of (15)N NMR relaxation data using the extended model free formalism reveals that CBM13 tumbles as an oblate ellipsoid (D( parallel)/D( perpendicular) = 0.80 +/- 0.02) and that its backbone is relatively rigid on the sub-nanosecond time scale. In particular, the three binding sites show no distinct patterns of increased internal mobility. Ligand-induced chemical shift changes in the (1)H-(15)N HSQC spectra of CBM13 were monitored as a function of increasing concentrations of L-arabinose, lactose, D-xylose, xylobiose, xylotetraose, and xylohexaose. Patterns of shift perturbations for well-reso...Continue Reading

References

Sep 1, 1995·Journal of Biomolecular NMR·D S WishartB D Sykes
Dec 2, 2000·Journal of Biomolecular NMR·D Sharma, K Rajarathnam

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Citations

Mar 22, 2006·Extremophiles : Life Under Extreme Conditions·Jörg Kleine, Wolfgang Liebl
Oct 22, 2003·Current Opinion in Structural Biology·Heide KogelbergJesús Jiménez-Barbero
Jul 2, 2003·Current Opinion in Microbiology·Dalia Shallom, Yuval Shoham
May 4, 2004·Journal of Biotechnology·Shigeyasu ItoTsunemi Hasegawa
Dec 12, 2013·Journal of Molecular Biology·Julie M GrondinSteven P Smith
Sep 11, 2012·Journal of Structural Biology·Daohua JiangXuejun C Zhang
Mar 9, 2004·The Journal of Biological Chemistry·Joanna L HenshawHarry J Gilbert
Jun 25, 2004·The Biochemical Journal·Alisdair B BorastonGideon J Davies
Apr 8, 2017·Bioscience, Biotechnology, and Biochemistry·Ayako MikiMasayuki Oda

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