%0 Journal Article
%A Ohlin, Mats
%A von Schantz, Laura
%A Schrader, Tobias E.
%A Ostermann, Andreas
%A Logan, Derek T.
%A Fisher, S. Zoë
%T Crystallization, neutron data collection, initial structure refinement and analysis of a xyloglucan heptamer bound to an engineered carbohydrate-binding module from xylanase
%J Acta crystallographica / F
%V 71
%N 8
%@ 2053-230X
%C Oxford [u.a.]
%I Blackwell
%M FZJ-2015-06689
%P 1072 - 1077
%D 2015
%X Carbohydrate-binding modules (CBMs) are discrete parts of carbohydrate-hydrolyzing enzymes that bind specific types of carbohydrates. Ultra high-resolution X-ray crystallographic studies of CBMs have helped to decipher the basis for specificity in carbohydrate-protein interactions. However, additional studies are needed to better understand which structural determinants confer which carbohydrate-binding properties. To address these issues, neutron crystallographic studies were initiated on one experimentally engineered CBM derived from a xylanase, X-2 L110F, a protein that is able to bind several different plant carbohydrates such as xylan, [beta]-glucan and xyloglucan. This protein evolved from a CBM present in xylanase Xyn10A of Rhodothermus marinus. The protein was complexed with a branched xyloglucan hepta­saccharide. Large single crystals of hydrogenous protein (~1.6 mm3) were grown at room temperature and subjected to H/D exchange. Both neutron and X-ray diffraction data sets were collected to 1.6 Å resolution. Joint neutron and X-ray refinement using phenix.refine showed significant density for residues involved in carbohydrate binding and revealed the details of a hydrogen-bonded water network around the binding site. This is the first report of a neutron structure of a CBM and will add to the understanding of protein-carbohydrate binding interactions.
%F PUB:(DE-HGF)16
%9 Journal Article
%U <Go to ISI:>//WOS:000359352700026
%$ pmid:26249702
%R 10.1107/S2053230X15011383
%U https://juser.fz-juelich.de/record/276224