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@ARTICLE{Samygina:81600,
author = {Samygina, V. R. and Moiseev, V. M. and Rodina, E. V. and
Vorobyeva, N. N. and Popov, A. N. and Kurilova, S. A. and
Nazarova, T. I. and Avaeva, S. M. and Bartunik, H. D. and
DESY},
title = {{R}eversible inhibition of {E}scherichia coli inorganic
pyrophosphatase by fluoride: trapped catalytic intermediates
in cryo-crystallographic studies},
journal = {Journal of molecular biology},
volume = {366},
issn = {0022-2836},
address = {Amsterdam [u.a.]},
publisher = {Elsevier},
reportid = {PHPPUBDB-6428},
pages = {1305-1317},
year = {2007},
abstract = {Here, we describe high-resolution X-ray structures of
Escherichia coli inorganic pyrophosphatase (E-PPase)
complexed with the substrate, magnesium, or manganese
pyrophosphate. The structures correspond to steps in the
catalytic synthesis of enzyme-bound pyrophosphate (PP(i)) in
the presence of fluoride as an inhibitor of hydrolysis. The
catalytic reaction intermediates were trapped applying a new
method that we developed for initiating hydrolytic activity
in the E-PPase crystal. X-ray structures were obtained for
three consecutive states of the enzyme in the course of
hydrolysis. Comparative analysis of these structures showed
that the Mn2+-supported hydrolysis of the phosphoanhydride
bond is followed by a fast release of the leaving phosphate
from the P1 site. The electrophilic phosphate P2 is trapped
in the "down" conformation. Its movement into the "up"
position most likely represents the rate-limiting step of
Mn2+-supported hydrolysis. We further determined the crystal
structure of the Arg43Gln mutant variant of E-PPase
complexed with one phosphate and four Mn ions.},
keywords = {Binding Sites / Catalysis / Diphosphates: chemistry /
Diphosphates: pharmacology / Enzyme Activation / Escherichia
coli: enzymology / Fluorides: chemistry / Fluorides:
pharmacology / Hydrogen-Ion Concentration / Hydrolysis /
Inorganic Pyrophosphatase: chemistry / Inorganic
Pyrophosphatase: genetics / Inorganic Pyrophosphatase:
metabolism / Magnesium: chemistry / Magnesium: pharmacology
/ Manganese: chemistry / Manganese: pharmacology / Models,
Molecular / Mutation / Protein Isoforms / Substrate
Specificity / X-Ray Diffraction: methods / Diphosphates (NLM
Chemicals) / Fluorides (NLM Chemicals) / Protein Isoforms
(NLM Chemicals) / Magnesium (NLM Chemicals) / Manganese (NLM
Chemicals) / Inorganic Pyrophosphatase (NLM Chemicals)},
cin = {MPG},
ddc = {570},
cid = {$I:(DE-H253)MPG_-2012_-20120307$},
pnm = {DORIS Beamline BW6 (POF1-550)},
pid = {G:(DE-H253)POF1-BW6-20130405},
experiment = {EXP:(DE-H253)D-BW6-20150101},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:17196979},
UT = {WOS:000244649500021},
doi = {10.1016/j.jmb.2006.11.082},
url = {https://bib-pubdb1.desy.de/record/81600},
}