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PDBsum entry 1e47
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Aldolase (class ii)
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PDB id
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1e47
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Contents |
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* Residue conservation analysis
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References listed in PDB file
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Key reference
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Title
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Structures of l-Fuculose-1-Phosphate aldolase mutants outlining motions during catalysis.
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Authors
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A.C.Joerger,
C.Mueller-Dieckmann,
G.E.Schulz.
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Ref.
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J Mol Biol, 2000,
303,
531-543.
[DOI no: ]
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PubMed id
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Abstract
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The crystal structures of l-fuculose-1-phosphate aldolase (FucA) with and
without a ligated analogue of dihydroxyacetone phosphate (DHAP) and of a number
of active center mutants have resulted in a model of the catalytic mechanism.
This model has now been confirmed by structural analyses of further mutations at
the zinc coordination sphere and at the phosphate site. In addition, these
mutants have revealed new aspects of the catalysis: the hydroxyl group of
Tyr113' (from a neighboring subunit), which sits just outside the zinc
coordination sphere, steers DHAP towards a productive binding mode at the zinc
ion; Glu73 contacts zinc in between the two ligand positions intended for the
DHAP oxygen atoms and thus avoids blocking of these positions by a tetrahedrally
coordinated hydroxy ion; the FucA polypeptide does not assume its minimum energy
state but oscillates between two states of elevated energy as demonstrated by a
mutant in a minimum energy state. The back and forth motion involves a mobile
loop connecting the phosphate site with intersubunit motions and thus with the
Brownian motion of the solvent. The phosphate group is bound strongly at a given
distance to the zinc ion, which prevents the formation of too tight a DHAP:zinc
complex. This observation explains our failure to find mutants that accept
phosphate-free substitutes for DHAP. The FucA zinc coordination sphere is
compared with that of carbonic anhydrase.
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Figure 2.
Figure 2. Stereo views of the active center of FucA: (a)
the complex FucA:Sulf with a sulfate ion from the
crystallization buffer bound at the phosphate site [Dreyer and
Schulz 1993 and Dreyer and Schulz 1996a] containing the mobile
loop (23-27) (orange); (b) the complex FucA:PGH with the
transition state analogue phosphoglycolohydroxamate [Dreyer and
Schulz 1996b] showing the solidified loop (23-27) and including
a model (orange) of bound Image -lactaldehyde and solidified
Tyr209' [Joerger et al 2000].
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Figure 7.
Figure 7. The zinc coordination spheres of FucA:Sulf
(yellow), FucA:PGH (red), FucA mutants Glu73->Ser (blue),
Glu73->Gln (light blue/grey), E73Q/Y113F/Y209F (magenta),
Ser71->Gln (green) and of carbonic anhydrase (pink, tetrahedron
drawn out) [Hakansson et al 1992]. The structures were
superimposed on the contacting histidine nitrogen atoms and the
zinc ion. The N  epsilon,
Greek atoms of histidine 92, 94 and 155 and the zinc ion of FucA
are shown in the respective colors. They correspond to His96-N
epsilon,
Greek , His94-N epsilon,
Greek , and His119-Nd and the zinc ion of carbonic anhydrase,
respectively.
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The above figures are
reprinted
by permission from Elsevier:
J Mol Biol
(2000,
303,
531-543)
copyright 2000.
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Secondary reference #1
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Title
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Catalytic action of fuculose 1-Phosphate aldolase (class ii) as derived from structure-Directed mutagenesis.
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Authors
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A.C.Joerger,
C.Gosse,
W.D.Fessner,
G.E.Schulz.
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Ref.
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Biochemistry, 2000,
39,
6033-6041.
[DOI no: ]
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PubMed id
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Secondary reference #2
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Title
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Catalytic mechanism of the metal-Dependent fuculose aldolase from escherichia coli as derived from the structure.
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Authors
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M.K.Dreyer,
G.E.Schulz.
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Ref.
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J Mol Biol, 1996,
259,
458-466.
[DOI no: ]
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PubMed id
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Figure 3.
Figure 3. Stereo view of the active center of FucA with positive (green) and negative (red) difference density at 23.5s
levels after soaking with PGH. Phases were taken from the unligated structure without sulfate, chloride and water. The
depicted residues are 21 to 31, 40 to 46 and 70 to 162 (black). Residues of the neighboring subunit are 109' to 123', 159'
to 174' and 198' to 206' (blue). The fitted PGH model is given in gold. The side-chains of Ala27, Asn29, Thr43, Ser71,
Ser72, Glu73 (still in the Zn
2+
coordination sphere), Phe131, Tyr113' and Phe206' are introduced.
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Figure 5.
Figure 5. B-factor distribution of FucA in crystal form K (continuous line). The dotted line depicts the B-factor
distribution of the complex FucA:PGH.
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The above figures are
reproduced from the cited reference
with permission from Elsevier
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Secondary reference #3
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Title
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The spatial structure of the class ii l-Fuculose-1-Phosphate aldolase from escherichia coli.
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Authors
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M.K.Dreyer,
G.E.Schulz.
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Ref.
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J Mol Biol, 1993,
231,
549-553.
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PubMed id
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