Báo cáo khoa học: Determination of thioxylo-oligosaccharide binding to family 11 xylanases using electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry and X-ray crystallography pot
... Calcium and peptide binding to folded and
unfolded conformations of cardiac troponin C. Electro-
spray ionization and Fourier transform ion cyclotron
resonance mass spectrometry. Eur J Mass Spectrom ... al. Thioxylo-oligosaccharide binding to xylanases
FEBS Journal 272 (2005) 2317–2333 ª 2005 FEBS 2333
Determination of thioxylo-oligosaccharide bindi...
... the
absence of cofactor reduction.
Stopped-flow kinetic studies
Reduction of the FAD/NADPH domain of NR1 was
investigated by stopped-flow methods using a photodiode
array detector. Aggregation of the ... units. To
enable accurate determination of the FAD potentials,
titrations of the NR1 FAD/NADPH domain were per-
formed on samples of identical concentration over small
range...
... to
water and couples the free energy of this reaction to
the generation of a transmembrane proton gradient.
The mitochondrial enzyme consists of up to 13 subun-
its, of which only the key subunits ... group of heme a, and most likely presents the
cofactor in a spatial orientation suitable for optimal transfer to its target
site within subunit I of cytochrome c oxidase....
... concen-
tration was determined using an e
260 nm
value of
7.0 · 10
3
m
)1
Æcm
)1
.
Expression and purification of Mhr1
For the expression of recombinant Mhr1, E. coli BL21(DE3)
pLysS DrecA cells were transformed ... recombina-
tion of mtDNA in Saccharomyces cerevisiae. The
mhr1-1 mutation causes defects in mtDNA duplication,
partitioning to bud, and recovery of homoplasmy, al...
... FEBS
Characterization of HbpR binding by site-directed
mutagenesis of its DNA -binding site and by deletion
of the effector domain
David Tropel
1
* and Jan R. van der Meer
1,2
1 Process of Environmental ... the HbpR–DNA binding interactions were investigated
by site-directed mutagenesis of the operator region and by DNA -binding
assays using purified HbpR. Mutations tha...
... denaturation and renatur-
ation manipulations, the fast annealing (fast renaturation)
and slow annealing (slow renaturation) procedures
(Materials and methods) were performed after the primer
2 and ... primer 2 and primer 3 under fast
and slow annealing conditions. (A) Primer extension with primer 2
under slow and fast annealing conditions: lanes 1 and 2, primer
extension prod...
...
of syntactic analysis of Estonian using
Constraint Grammar. Constraint Gram-
mar framework divides parsing into two
different modules: morphological disam-
biguation and determination of ... The process
of syntactic analysis consists of three stages: mor-
phological disambiguation, identification of clause
boundaries, and identification of syntactic func-
tions o...
... human
glioblastoma cell migration into the brain through modifications
to the actin cytoskeleton and levels of expression of small
GTPases. J. Neuropathol. Exp. Neurol. 61, 585–596.
134 S. Andre
´
et ... ligand properties of the
neoglycoproteins, we determined their binding to cells and
to tissue sections as well as their biodistribution in mice after
injection of radioiodi...
... substrate
binding site and the role of the metal cofactor in the
enzyme.
Results
Cloning, expression and purification of BphH
The bphH gene was inserted into the expression vectors
pT7-7 [11] , pEMBL18 ... values
of 6.0 and 8.0 for catalysis.
Number of metal ion bound per enzyme and
determination of coordination number
The equilibrium concentration of Mg
2+
bound to...
... addition of 975 lLofNa
2
CO
3
(50 m
M
). One
unit (U) of activity was defined a s the quantity of enzyme
necessary to release 1 lmol of 2-chloro-4-nitrophenol per
min under the assay conditions, using ... concerning the
conservation of the catalytic machinery [8].
Evolutionary relationships
In order to draw the present-day evolutionary picture of the
family GH-57, several e...