 |
|
|
|
|
|
|
|
|
|
|
 |
|
|
|
|
|
|
| |
|
|
| |
|
DOI no:
|
Biochemistry
36:16032-16039
(1997)
|
|
PubMed id:
|
|
|
|
|
|
| |
|
The Streptomyces lividans family 12 endoglucanase: construction of the catalytic cre, expression, and X-ray structure at 1.75 A resolution.
|
|
G.Sulzenbacher,
F.Shareck,
R.Morosoli,
C.Dupont,
G.J.Davies.
|
|
|
|
|
| |
ABSTRACT
|
|
|
|
| |
|
|
Cellulases are the glycoside hydrolases responsible for the enzymatic breakdown
of the structural plant polymer cellulose. Together with xylanases they
counteract the lmitless accumulation of plant biomass in nature and are of
considerable fundamental and biotechnological interest. Endoglucanase CelB from
Streptomyces lividans performs hydrolysis of the beta-1,4-glycosidic bonds of
cellulose, with net retention of anomeric configuration. The enzyme is a member
of glycoside hydrolase family 12 [Henrissat, B., and Bairoch, A. (1996) Biochem.
J. 316, 695-696], which had previously eluded detailed structural analysis. A
truncated, but cataytically competent form of CelB, locking the flexible linker
region and cellulose-binding domain, has been constructed and overexpressed in a
S. lividans expression system. The three-dimensional X-ray structure of the
resulting catalytic domain, CelB2, has been solved by conventional multiple
isomorphous replacement methods and refined to an R factor of 0.187 at 1.75 A
resolution. The overall fold of the enzyme shows a remarkable similarity to that
of family 11 xylanases, as previously predicted by hydrophobic clustering
analysis [Törrönen, A., Kubicek, C.P., and Henrissat, B. (1993) FEBS Lett.
321, 135-139]. The 23 kDa protein presents a jelly-roll topology, built up
mainly by antiparallel beta-sheets arranged in a sandwich-like manner. A deep
substrate-binding cleft runs across the surface, as has been observed in other
endoglucanase structures, and is potentially able to accommodate up to five
binding subsites. The likely catalytic nucleophile and Brønsted acid/base,
residues Glu 120 and Glue 203, respectively, have their carboxylate groups
separated by a distance of approximately 7.0 A and are located approximately 15
A from one end of the cleft, implying a -3 to +2 active site.
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
 |
 |
 |
Literature references that cite this PDB file's
key reference
|
|
| |
PubMed id
|
|
Reference
|
|
|
|
|
|
T.M.Gloster,
F.M.Ibatullin,
K.Macauley,
J.M.Eklöf,
S.Roberts,
J.P.Turkenburg,
M.E.Bjørnvad,
P.L.Jørgensen,
S.Danielsen,
K.S.Johansen,
T.V.Borchert,
K.S.Wilson,
H.Brumer,
and
G.J.Davies
(2007).
Characterization and three-dimensional structures of two distinct bacterial xyloglucanases from families GH5 and GH12.
|
| |
J Biol Chem, 282,
19177-19189.
|
|
|
PDB codes:
|
|
|
|
|
|
|
|
G.Sulzenbacher,
C.Bignon,
T.Nishimura,
C.A.Tarling,
S.G.Withers,
B.Henrissat,
and
Y.Bourne
(2004).
Crystal structure of Thermotoga maritima alpha-L-fucosidase. Insights into the catalytic mechanism and the molecular basis for fucosidosis.
|
| |
J Biol Chem, 279,
13119-13128.
|
|
|
PDB codes:
|
|
|
|
|
|
|
|
L.Hildén,
and
G.Johansson
(2004).
Recent developments on cellulases and carbohydrate-binding modules with cellulose affinity.
|
| |
Biotechnol Lett, 26,
1683-1693.
|
|
|
|
|
|
|
M.Sandgren,
P.J.Gualfetti,
A.Shaw,
L.S.Gross,
M.Saldajeno,
A.G.Day,
T.A.Jones,
and
C.Mitchinson
(2003).
Comparison of family 12 glycoside hydrolases and recruited substitutions important for thermal stability.
|
| |
Protein Sci, 12,
848-860.
|
|
|
PDB codes:
|
|
|
|
|
|
|
|
M.Sandgren,
P.J.Gualfetti,
C.Paech,
S.Paech,
A.Shaw,
L.S.Gross,
M.Saldajeno,
G.I.Berglund,
T.A.Jones,
and
C.Mitchinson
(2003).
The Humicola grisea Cel12A enzyme structure at 1.2 A resolution and the impact of its free cysteine residues on thermal stability.
|
| |
Protein Sci, 12,
2782-2793.
|
|
|
PDB codes:
|
|
|
|
|
|
|
|
S.Khademi,
D.Zhang,
S.M.Swanson,
A.Wartenberg,
K.Witte,
and
E.F.Meyer
(2002).
Determination of the structure of an endoglucanase from Aspergillus niger and its mode of inhibition by palladium chloride.
|
| |
Acta Crystallogr D Biol Crystallogr, 58,
660-667.
|
|
|
PDB codes:
|
|
|
|
|
|
|
|
S.Zhang,
D.C.Irwin,
and
D.B.Wilson
(2000).
Site-directed mutation of noncatalytic residues of Thermobifida fusca exocellulase Cel6B.
|
| |
Eur J Biochem, 267,
3101-3115.
|
|
|
|
|
|
|
G.Henriksson,
A.Nutt,
H.Henriksson,
B.Pettersson,
J.Ståhlberg,
G.Johansson,
and
G.Pettersson
(1999).
Endoglucanase 28 (Cel12A), a new Phanerochaete chrysosporium cellulase.
|
| |
Eur J Biochem, 259,
88-95.
|
|
|
|
|
|
|
G.Sulzenbacher,
L.F.Mackenzie,
K.S.Wilson,
S.G.Withers,
C.Dupont,
and
G.J.Davies
(1999).
The crystal structure of a 2-fluorocellotriosyl complex of the Streptomyces lividans endoglucanase CelB2 at 1.2 A resolution.
|
| |
Biochemistry, 38,
4826-4833.
|
|
|
PDB code:
|
|
|
|
|
|
|
|
M.W.Bauer,
L.E.Driskill,
W.Callen,
M.A.Snead,
E.J.Mathur,
and
R.M.Kelly
(1999).
An endoglucanase, EglA, from the hyperthermophilic archaeon Pyrococcus furiosus hydrolyzes beta-1,4 bonds in mixed-linkage (1-->3),(1-->4)-beta-D-glucans and cellulose.
|
| |
J Bacteriol, 181,
284-290.
|
|
|
|
|
|
|
S.Kawaminami,
H.Takahashi,
S.Ito,
Y.Arata,
and
I.Shimada
(1999).
A multinuclear NMR study of the active site of an endoglucanase from a strain of Bacillus. Use of Trp residues as structural probes.
|
| |
J Biol Chem, 274,
19823-19828.
|
|
|
|
|
|
The most recent references are shown first.
Citation data come partly from CiteXplore and partly
from an automated harvesting procedure. Note that this is likely to be
only a partial list as not all journals are covered by
either method. However, we are continually building up the citation data
so more and more references will be included with time.
Where a reference describes a PDB structure, the PDB
codes are
shown on the right.
|
222 a.a.
