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@ARTICLE{Kachalova:94652,
author = {Kachalova, G. and Decker, K. and Holt, A. and Bartunik, H.
D. and DESY},
title = {{C}rystallographic snapshots of the complete reaction cycle
of nicotine degradation by an amine oxidase of the monoamine
oxidase ({MAO}) family},
journal = {Proceedings of the National Academy of Sciences of the
United States of America},
volume = {108},
issn = {1091-6490},
address = {Washington, DC},
publisher = {Academy},
reportid = {PHPPUBDB-18289},
pages = {4800-4805},
year = {2011},
abstract = {FAD-linked oxidases constitute a class of enzymes which
catalyze dehydrogenation as a fundamental biochemical
reaction, followed by reoxidation of reduced flavin. Here,
we present high-resolution crystal structures showing the
flavoenzyme 6-hydroxy-l-nicotine oxidase in action. This
enzyme was trapped during catalytic degradation of the
native substrate in a sequence of discrete reaction states
corresponding to the substrate-reduced enzyme, a complex of
the enzyme with the intermediate enamine product and
formation of the final aminoketone product. The inactive
d-stereoisomer binds in mirror symmetry with respect to the
catalytic axis, revealing absolute stereospecificity of
hydrogen transfer to the flavin. The structural data suggest
deprotonation of the substrate when bound at the active
site, an overall binary complex mechanism and oxidation by
direct hydride transfer. The amine nitrogen has a critical
role in the dehydrogenation step and may activate
carbocation formation at the α-carbon via delocalization
from the lone pair to σ* C(α)-H. Enzymatically assisted
hydrolysis of the intermediate product occurs at a remote
(P site) cavity. Substrate entry and product exit follow
different paths. Structural and kinetic data suggest that
substrate can also bind to the reduced enzyme, associated
with slower reoxidation as compared to the rate of
reoxidation of free enzyme. The results are of general
relevance for the mechanisms of flavin amine oxidases.},
keywords = {Arthrobacter: enzymology / Bacterial Proteins: chemistry /
Catalytic Domain / Crystallography, X-Ray / Kinetics /
Monoamine Oxidase: chemistry / Nicotine: chemistry /
Oxidation-Reduction / Oxidoreductases Acting on CH-NH Group
Donors: chemistry / Structure-Activity Relationship /
Bacterial Proteins (NLM Chemicals) / Nicotine (NLM
Chemicals) / Monoamine Oxidase (NLM Chemicals) /
Oxidoreductases Acting on CH-NH Group Donors (NLM Chemicals)
/ 6-hydroxy-L-nicotine oxidase (NLM Chemicals)},
cin = {HASYLAB / MPG},
ddc = {000},
cid = {$I:(DE-H253)HASYLAB_-2012_-20130307$ /
$I:(DE-H253)MPG_-2012_-20120307$},
pnm = {DORIS Beamline BW6 (POF2-54G13)},
pid = {G:(DE-H253)POF2-BW6-20130405},
experiment = {EXP:(DE-H253)D-BW6-20150101},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:21383134},
pmc = {pmc:PMC3064382},
UT = {WOS:000288712200025},
doi = {10.1073/pnas.1016684108},
url = {https://bib-pubdb1.desy.de/record/94652},
}