Crystallization, neutron data collection, initial structure refinement and analysis of a xyloglucan heptamer bound to an engineered carbohydrate-binding module from xylanase.
Ohlin, Mats LU
; von Schantz, Laura
LU
; Schrader, Tobias E
; Ostermann, Andreas
; Logan, Derek
LU
and Fisher, S Zoë
(2015)
In Acta crystallographica. Section F, Structural biology communications
71(Pt 8).
p.1072-1077
- Abstract
- 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, β-glucan and xyloglucan. This protein evolved from a CBM present in xylanase Xyn10A of Rhodothermus... (More)
- 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, β-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 mm(3)) 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. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/7844491
- author
- Ohlin, Mats
LU
; von Schantz, Laura
LU
; Schrader, Tobias E
; Ostermann, Andreas
; Logan, Derek
LU
and Fisher, S Zoë
- organization
- publishing date
- 2015
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Acta crystallographica. Section F, Structural biology communications
- volume
- 71
- issue
- Pt 8
- pages
- 1072 - 1077
- publisher
- Wiley-Blackwell
- external identifiers
-
- pmid:26249702
- wos:000359352700026
- scopus:84938863810
- pmid:26249702
- ISSN
- 2053-230X
- DOI
- 10.1107/S2053230X15011383
- project
- Designed carbohydrate binding modules and molecular probes
- language
- English
- LU publication?
- yes
- id
- a9f4959f-911b-4810-b748-1feccf7a9922 (old id 7844491)
- date added to LUP
- 2016-04-01 14:25:34
- date last changed
- 2022-01-28 00:35:15
@article{a9f4959f-911b-4810-b748-1feccf7a9922,
abstract = {{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, β-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 mm(3)) 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.}},
author = {{Ohlin, Mats and von Schantz, Laura and Schrader, Tobias E and Ostermann, Andreas and Logan, Derek and Fisher, S Zoë}},
issn = {{2053-230X}},
language = {{eng}},
number = {{Pt 8}},
pages = {{1072--1077}},
publisher = {{Wiley-Blackwell}},
series = {{Acta crystallographica. Section F, Structural biology communications}},
title = {{Crystallization, neutron data collection, initial structure refinement and analysis of a xyloglucan heptamer bound to an engineered carbohydrate-binding module from xylanase.}},
url = {{http://dx.doi.org/10.1107/S2053230X15011383}},
doi = {{10.1107/S2053230X15011383}},
volume = {{71}},
year = {{2015}},
}