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@ARTICLE{Li:859977,
author = {Li, Zhihong and Zhang, Xiaoshuai and Wang, Qingqing and Li,
Chunran and Zhang, Nianying and Zhang, Xinkai and Xu, Birui
and Ma, Baoliang and Schrader, Tobias E. and Coates,
Leighton and Kovalevsky, Andrey and Huang, Yandong and Wan,
Qun},
title = {{U}nderstanding the p{H}-{D}ependent {R}eaction {M}echanism
of a {G}lycoside {H}ydrolase {U}sing {H}igh-{R}esolution
{X}-ray and {N}eutron {C}rystallographyülcih},
journal = {ACS catalysis},
volume = {8},
number = {9},
issn = {2155-5435},
address = {Washington, DC},
publisher = {ACS},
reportid = {FZJ-2019-00783},
pages = {8058 - 8069},
year = {2018},
abstract = {Glycoside hydrolases (GHs) commonly use the retaining or
inverting mechanisms to hydrolyze carbohydrates, and the
rates of catalysis are usually pH dependent. Deeper
understanding of these pH-dependent reaction mechanisms is
of great importance for protein engineering and drug design.
We used high-resolution X-ray crystallography to analyze the
sugar ring configurations of an oligosaccharide ligand
during hydrolysis for the family 11 GH, and the results
support the 1S3 → 4H3 → 4C1 conformational itinerary.
These results indicate that sugar ring flexibility may help
to distort and break the glycosidic bond. Constant pH
molecular dynamics simulations and neutron crystallography
demonstrate that the catalytic glutamate residue (E177) has
alternate conformational changes to transfer a proton to
cleave the glycosidic bond. Furthermore, a neutron
crystallography analysis shows that the H-bond length
between E177 and its nearby tyrosine residue (Y88) is
shortened when the pH increases, preventing E177 from
rotating downward and obtaining a proton from the solvent
for catalysis. This result indicates that the H-bond length
variation may play a key role in the pH-dependent reaction
mechanism. In summary, our results demonstrate that both
sugar ring flexibility and protein dynamics are important in
the pH-dependent reaction mechanism and may help to engineer
GHs with different pH optima.},
cin = {JCNS-FRM-II / JCNS-1},
ddc = {540},
cid = {I:(DE-Juel1)JCNS-FRM-II-20110218 /
I:(DE-Juel1)JCNS-1-20110106},
pnm = {6G15 - FRM II / MLZ (POF3-6G15) / 6G4 - Jülich Centre for
Neutron Research (JCNS) (POF3-623) / 6215 - Soft Matter,
Health and Life Sciences (POF3-621)},
pid = {G:(DE-HGF)POF3-6G15 / G:(DE-HGF)POF3-6G4 /
G:(DE-HGF)POF3-6215},
experiment = {EXP:(DE-MLZ)BIODIFF-20140101},
typ = {PUB:(DE-HGF)16},
UT = {WOS:000444364800034},
doi = {10.1021/acscatal.8b01472},
url = {https://juser.fz-juelich.de/record/859977},
}