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PDBsum entry 1cxh
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Glycosyltransferase
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PDB id
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1cxh
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References listed in PDB file
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Key reference
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Title
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Crystallographic studies of the interaction of cyclodextrin glycosyltransferase from bacillus circulans strain 251 with natural substrates and products.
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Authors
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R.M.Knegtel,
B.Strokopytov,
D.Penninga,
O.G.Faber,
H.J.Rozeboom,
K.H.Kalk,
L.Dijkhuizen,
B.W.Dijkstra.
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Ref.
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J Biol Chem, 1995,
270,
29256-29264.
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PubMed id
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Note In the PDB file this reference is
annotated as "TO BE PUBLISHED".
The citation details given above were identified by an automated
search of PubMed on title and author
names, giving a
percentage match of
96%.
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Abstract
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Asp-229, Glu-257, and Asp-328 constitute the catalytic residues in cyclodextrin
glycosyl transferase from Bacillus circulans strain 251. Via site-directed
mutagenesis constructed D229N, E257Q, and D328N mutant proteins showed a
4,000-60,000-fold reduction of cyclization activity. A D229N/E257Q double mutant
showed a 700,000-fold reduction and was crystallized for use in soaking
experiments with alpha-cyclodextrin. Crystal structures were determined of wild
type CGTase soaked at elevated pH with alpha-cyclodextrin (resolution, 2.1 A)
and maltoheptaose (2.4 A). In addition, structures at cryogenic temperature were
solved of the unliganded enzyme (2.2 A) and of the D229N/E257Q mutant after
soaking with alpha-cyclodextrin (2.6 A). In the crystals soaked in
alpha-cyclodextrin and maltoheptaose, a maltotetraose molecule is observed to
bind in the active site. Residue 229 is at hydrogen bonding distance from the
C-6 hydroxyl group of the sugar, which after cleavage will contain the new
reducing end. In the D229N/E257Q double mutant structure, two
alpha-cyclodextrins are observed to replace two maltoses at the E-domain, thus
providing structural information on product inhibition via binding to the
enzyme's raw starch binding domain.
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Secondary reference #1
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Title
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X-Ray structure of cyclodextrin glycosyltransferase complexed with acarbose. Implications for the catalytic mechanism of glycosidases.
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Authors
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B.Strokopytov,
D.Penninga,
H.J.Rozeboom,
K.H.Kalk,
L.Dijkhuizen,
B.W.Dijkstra.
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Ref.
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Biochemistry, 1995,
34,
2234-2240.
[DOI no: ]
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PubMed id
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Secondary reference #2
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Title
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Nucleotide sequence and X-Ray structure of cyclodextrin glycosyltransferase from bacillus circulans strain 251 in a maltose-Dependent crystal form.
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Authors
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C.L.Lawson,
R.Van montfort,
B.Strokopytov,
H.J.Rozeboom,
K.H.Kalk,
G.E.De vries,
D.Penninga,
L.Dijkhuizen,
B.W.Dijkstra.
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Ref.
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J Mol Biol, 1994,
236,
590-600.
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PubMed id
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Secondary reference #3
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Title
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Maltodextrin-Dependent crystallization of cyclomaltodextrin glucanotransferase from bacillus circulans.
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Authors
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C.L.Lawson,
J.Bergsma,
P.M.Bruinenberg,
G.De vries,
L.Dijkhuizen,
B.W.Dijkstra.
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Ref.
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J Mol Biol, 1990,
214,
807-809.
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PubMed id
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