... oligopeptide sequences and using PCR, the
gene of the cyclodextrinase from Flavobacterium sp. no. 92
wasderivedfromthegenomicDNA.Thegenewas
sequenced and expressed in Escherichia coli; the gene pro-
duct ... longer
and more complex than the loops at the opposite barrel end.
The lengths of the b-strands in the barrel vary from two (b14)
to seven (b11) resi...
... II. The hood domain is mainly built
up by residues from the C-terminus and consists of a
two-stranded anti-parallel b-sheet composed of b2 and
b9 and an a-helix from the C-terminus (Fig. 1A). The
two ... atoms,
whereas the side chain of Lys173 interacts with both
O6 and N7 (Fig. 1C). The 2¢OH of the ribose is hydro-
gen bonded to the main chain and the...
... [57].
Isolation and cloning of the SynMGST
and E.coliMGST
The coding sequence for the SynMGST, corresponding to
the complementary strand of the nucleotide sequence from
89 254 to 89 667 in the Synechocystis ... above and in
[1]). We investigated representative gene products from
the E. coli and Synechocystis sp. bacteria further, to
gain insight into the funct...
... GSH and
to the diol, mono-acetate and di-acetate forms of the
parent drug, respectively. The latter three species reflect the
hydrolysis of the compound in solution (see the results
below with the ... reactivity of S23906-1 towards the
bionucleophiles DNA and GSH. The covalent binding of
the drug to GSH reduced the formation of DNA adducts
and slightly...
... distortion of the )1
sugar and by increasing the pK
a
of the catalytic acid. The
hydroxyl group of Tyr214 makes a major contribution to
the positioning of the N-acetyl group of the )1sugar.Taken
together, ... simplicity, and in line with the results from the other calculations, the <0.0 value is allocated to Asp140
and > 20.0 to Asp142.
b
The calc...
... 7).
Although the spectral characteristics of the lac1 protein
suggest the absence of a type 1 copper site, this is not borne
out by analysis of the deduced amino acid sequence of the
enzyme. Thus, the ... amino acid sequence from
Ala25 to Asp41 corresponded to the N-terminal sequence of
the purifiedprotein,andtheputativepresequenceof 24amino
acids is a hydrophobic s...
... comprising the
N-terminal fragment and the first AAA-cassette of
PEX6 (PEX6N + D1–GAL4-BD). Neither of the cas-
settes alone nor the two AAA-cassettes together or the
N-terminal part together with the ... assigned their corresponding binding sites and elucidated the
importance of ATP-binding and -hydrolysis of Pex1p and Pex6p for their
interaction. We show that t...
... from comparison of the electronic absorption
and RR spectra of the wild type and Thr171Ser either
in the presence of the aromatic donor BHA or at
alkaline pH. Addition of saturating amounts of ... amounts of BHA
did not reveal any spectral differences between the wild
type and the mutant (data not shown). Furthermore,
the electronic absorption spectra of t...
... vector. The nucleotides are numbered
from the translation start site (ATG). The effect of the wild-type and mutants was measured as the mean of the firefly luciferase levels nor-
malized by the Renilla ... conjunction with the luciferase gene in pGL3-
basic vector. The nucleotides are numbered from the translation start site. The effect of the wild-type and...
... Stereo figures of the model of the bound substrate in the AD2 E526A mutant (A) and D524A mutant (B). The structures of bound
sugars and the side-chains of three acidic residues in the conserved ... causes the distortion and twist-
ing of the substrate, so that the glycosidic oxygen faces
towards the bottom of the deep cleft.
Enzyme–substrate interactions...