DB code: S00150

RLCP classification 1.30.36211.992 : Hydrolysis
CATH domain 2.60.120.180 : Jelly Rolls Catalytic domain
E.C. 3.2.1.4
CSA
M-CSA
MACiE

CATH domain Related DB codes (homologues)
2.60.120.180 : Jelly Rolls S00533 S00151 D00504 D00538

Uniprot Enzyme Name
UniprotKB Protein name Synonyms CAZy Pfam
O33897
Cellulase
EC 3.2.1.4
GH12 (Glycoside Hydrolase Family 12)
PF01670 (Glyco_hydro_12)
[Graphical View]
O00095
Endo-beta-1,4-glucanase (Beta-1,4-glucanase)
EC 3.2.1.4
GH12 (Glycoside Hydrolase Family 12)
PF01670 (Glyco_hydro_12)
[Graphical View]
Q8NJY3
Endoglucanase
GH12 (Glycoside Hydrolase Family 12)
PF01670 (Glyco_hydro_12)
[Graphical View]
Q8NJY6
Endoglucanase
GH12 (Glycoside Hydrolase Family 12)
PF01670 (Glyco_hydro_12)
[Graphical View]

KEGG enzyme name
cellulase
endo-1,4-beta-D-glucanase
beta-1,4-glucanase
beta-1,4-endoglucan hydrolase
celluase A
cellulosin AP
endoglucanase D
alkali cellulase
cellulase A 3
celludextrinase
9.5 cellulase
avicelase
pancellase SS
1,4-(1,3
1,4)-beta-D-glucan 4-glucanohydrolase

UniprotKB: Accession Number Entry name Activity Subunit Subcellular location Cofactor
O33897 O33897_RHOMR
O00095 O00095_TRIRE
Q8NJY3 Q8NJY3_9ASCO
Q8NJY6 Q8NJY6_9HYPO

KEGG Pathways
Map code Pathways E.C.
MAP00500 Starch and sucrose metabolism

Compound table
Substrates Products Intermediates
KEGG-id C00760 C00478 C00551 C00001 C00760 C00551
E.C.
Compound Cellulose Lichenin beta-D-Glucan H2O Cellulose beta-D-Glucan Transition-state for glycosylated enzyme Glycosylated enzyme intermediate
Type polysaccharide carbohydrate polysaccharide H2O polysaccharide polysaccharide
ChEBI 15377
PubChem 439241
46173706
22247451
962
46173706
1h0bA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound
1h0bB Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound
2bw8A Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound
2bw8B Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound
2bwaA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Bound:GLC-BGC-BGC Unbound
2bwaB Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Bound:GLC-BGC Unbound
2bwcA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Bound:GLC-BGC-BGC Unbound
2bwcB Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Bound:GLC-BGC-BGC-BGC Unbound Unbound Bound:BGC-BGC-BGC Unbound
1h8vA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound
1h8vB Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound
1h8vC Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound
1h8vD Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound
1h8vE Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound
1h8vF Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound
1oa2A Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound
1oa2B Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound
1oa2C Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound
1oa2D Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound
1oa2E Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound
1oa2F Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound
1olqA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound
1olqB Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound
1olrA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound
1uu4A Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Bound:BGC-BGC
1uu5A Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Bound:BGC-BGC-BGC-BGC Unbound
1uu6A Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Bound:BGC-BGC-BGC-BGC Unbound Unbound Unbound Unbound
1w2uA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Bound:BGC-BGC-BGC-BGC Unbound Unbound Unbound Unbound
1oa3A Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound
1oa3B Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound
1oa3C Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound
1oa3D Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain Unbound Unbound Unbound Unbound Unbound

Reference for Active-site residues
resource references E.C.
literature [2], [3], [5], [7]

Active-site residues
PDB Catalytic residues Cofactor-binding residues Modified residues Main-chain involved in catalysis Comment
1h0bA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 102;ASP 106;GLU 124;GLU 207
1h0bB Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 102;ASP 106;GLU 124;GLU 207
2bw8A Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 102;ASP 106;GLU 124;GLU 207
2bw8B Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 102;ASP 106;GLU 124;GLU 207
2bwaA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 102;ASP 106;GLU 124;GLU 207
2bwaB Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 102;ASP 106;GLU 124;GLU 207
2bwcA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 102;ASP 106;GLU 124;GLU 207
2bwcB Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 102;ASP 106;GLU 124;GLU 207
1h8vA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 95;ASP 99;GLU 116;GLU 200
1h8vB Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 95;ASP 99;GLU 116;GLU 200
1h8vC Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 95;ASP 99;GLU 116;GLU 200
1h8vD Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 95;ASP 99;GLU 116;GLU 200
1h8vE Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 95;ASP 99;GLU 116;GLU 200
1h8vF Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 95;ASP 99;GLU 116;GLU 200
1oa2A Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 95;ASP 99;GLU 116;GLU 200
1oa2B Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 95;ASP 99;GLU 116;GLU 200
1oa2C Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 95;ASP 99;GLU 116;GLU 200
1oa2D Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 95;ASP 99;GLU 116;GLU 200
1oa2E Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 95;ASP 99;GLU 116;GLU 200
1oa2F Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 95;ASP 99;GLU 116;GLU 200
1olqA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 95;ASP 99;GLU 116;GLU 200
1olqB Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 95;ASP 99;GLU 116;GLU 200
1olrA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 99;ASP 103;GLU 120;GLU 205
1uu4A Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 99;ASP 103;GLU 120;GLU 205
1uu5A Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 99;ASP 103;GLU 120;GLU 205
1uu6A Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 99;ASP 103;GLU 120;GLU 205
1w2uA Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 99;ASP 103;GLU 120;GLU 205
1oa3A Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 95;ASP 99;GLU 116;GLU 200
1oa3B Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 95;ASP 99;GLU 116;GLU 200
1oa3C Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 95;ASP 99;GLU 116;GLU 200
1oa3D Pdbj logo s Rasmollogo id Rasmollogo chain Mmcif id Mmcif chain ASN 95;ASP 99;GLU 116;GLU 200

References for Catalytic Mechanism
References Sections No. of steps in catalysis
[2]
p.16037-16038
[3]
Scheme 1, p.139-140
[4]
p.301
[6]
p.888-889
[9]
p.1510-1511
[11]
Fig.2, p.252-254
[12]
p.63-64, p.67

References
[1]
Resource
Comments
Medline ID
PubMed ID 8477842
Journal FEBS Lett
Year 1993
Volume 321
Pages 135-9
Authors Torronen A, Kubicek CP, Henrissat B
Title Amino acid sequence similarities between low molecular weight endo-1,4-beta-xylanases and family H cellulases revealed by clustering analysis.
Related PDB
Related UniProtKB
[2]
Resource
Comments
Medline ID
PubMed ID 9440876
Journal Biochemistry
Year 1997
Volume 36
Pages 16032-9
Authors Sulzenbacher G, Shareck F, Morosoli R, Dupont C, Davies GJ
Title The Streptomyces lividans family 12 endoglucanase: construction of the catalytic cre, expression, and X-ray structure at 1.75 A resolution.
Related PDB
Related UniProtKB
[3]
Resource
Comments
Medline ID
PubMed ID 9806895
Journal Biochem J
Year 1998
Volume 336
Pages 139-45
Authors Zechel DL, He S, Dupont C, Withers SG
Title Identification of Glu-120 as the catalytic nucleophile in Streptomyces lividans endoglucanase celB.
Related PDB
Related UniProtKB
[4]
Resource
Comments X-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS) OF 17-234.
Medline ID
PubMed ID 11327768
Journal J Mol Biol
Year 2001
Volume 308
Pages 295-310
Authors Sandgren M, Shaw A, Ropp TH, Wu S, Bott R, Cameron AD, Stahlberg J, Mitchinson C, Jones TA
Title The X-ray crystal structure of the Trichoderma reesei family 12 endoglucanase 3, Cel12A, at 1.9 A resolution.
Related PDB 1h8v
Related UniProtKB O00095
[5]
Resource
Comments
Medline ID
PubMed ID 12073090
Journal Curr Genet
Year 2002
Volume 41
Pages 89-98
Authors Goedegebuur F, Fowler T, Phillips J, van der Kley P, van Solingen P, Dankmeyer L, Power SD
Title Cloning and relational analysis of 15 novel fungal endoglucanases from family 12 glycosyl hydrolase.
Related PDB
Related UniProtKB
[6]
Resource
Comments X-RAY CRYSTALLOGRAPHY (1.8 ANGSTROMS) OF 38-260.
Medline ID
PubMed ID 12095262
Journal J Mol Biol
Year 2002
Volume 320
Pages 883-97
Authors Crennell SJ, Hreggvidsson GO, Nordberg Karlsson E
Title The structure of Rhodothermus marinus Cel12A, a highly thermostable family 12 endoglucanase, at 1.8 A resolution.
Related PDB 1h0b
Related UniProtKB O33897
[7]
Resource
Comments X-RAY CRYSTALLOGRAPHY (1.7 ANGSTROMS) OF 17-234.
Medline ID
PubMed ID 14627738
Journal Protein Sci
Year 2003
Volume 12
Pages 2782-93
Authors Sandgren M, Gualfetti PJ, Paech C, Paech S, Shaw A, Gross LS, Saldajeno M, Berglund GI, Jones TA, Mitchinson C
Title The Humicola grisea Cel12A enzyme structure at 1.2 A resolution and the impact of its free cysteine residues on thermal stability.
Related PDB 1olr 1olq
Related UniProtKB O00095 Q8NJY3
[8]
Resource
Comments X-RAY CRYSTALLOGRAPHY (1.5 ANGSTROMS) OF 17-234.
Medline ID
PubMed ID 12649442
Journal Protein Sci
Year 2003
Volume 12
Pages 848-60
Authors Sandgren M, Gualfetti PJ, Shaw A, Gross LS, Saldajeno M, Day AG, Jones TA, Mitchinson C
Title Comparison of family 12 glycoside hydrolases and recruited substitutions important for thermal stability.
Related PDB 1oa2 1oa3
Related UniProtKB O00095
[9]
Resource
Comments X-RAY CRYSTALLOGRAPHY (1.49 ANGSTROMS) OF 31-254.
Medline ID
PubMed ID 15364577
Journal J Mol Biol
Year 2004
Volume 342
Pages 1505-17
Authors Sandgren M, Berglund GI, Shaw A, Stahlberg J, Kenne L, Desmet T, Mitchinson C
Title Crystal complex structures reveal how substrate is bound in the -4 to the +2 binding sites of Humicola grisea Cel12A.
Related PDB 1uu4 1uu5 1uu6 1w2u
Related UniProtKB Q8NJY3 Q8NJY6
[10]
Resource
Comments
Medline ID
PubMed ID 15456402
Journal Biochem J
Year 2005
Volume 385
Pages 581-8
Authors Huang Y, Krauss G, Cottaz S, Driguez H, Lipps G
Title A highly acid-stable and thermostable endo-beta-glucanase from the thermoacidophilic archaeon Sulfolobus solfataricus.
Related PDB
Related UniProtKB
[11]
Resource
Comments
Medline ID
PubMed ID 15950056
Journal Prog Biophys Mol Biol
Year 2005
Volume 89
Pages 246-91
Authors Sandgren M, Stahlberg J, Mitchinson C
Title Structural and biochemical studies of GH family 12 cellulases: improved thermal stability, and ligand complexes.
Related PDB
Related UniProtKB
[12]
Resource
Comments X-ray crystallography
Medline ID
PubMed ID 16343530
Journal J Mol Biol
Year 2006
Volume 356
Pages 57-71
Authors Crennell SJ, Cook D, Minns A, Svergun D, Andersen RL, Nordberg Karlsson E
Title Dimerisation and an increase in active site aromatic groups as adaptations to high temperatures: X-ray solution scattering and substrate-bound crystal structures of Rhodothermus marinus endoglucanase Cel12A.
Related PDB 2bw8 2bwa 2bwc
Related UniProtKB

Comments
This family belongs to glycosidase family-12, which has a retaining mechanism.
The catalytic domain of this enzyme is homologous to those of another cellulase (S00533 in EzCatDB) and beta-1,4-Glucanase (D00504 in EzCatDB), whose catalytic mechanisms must be similar to that of this enzyme.
According to the literature [2], [3] & [7], the catalytic reaction proceeds as follows:
(1) Asn102 (of 1h0b) may modulate the activity of Glu207 as a acid-base.
(2) Glu207 (of 1h0b) acts as a general acid to protonate the leaving oxygen atom, leading to a oxocarbenium-like transtion state from cellulose substrate.
(3) pKa of Glu124 (of 1h0b), which acts as a nucleophile, is modulated by Asp106.
(4) Glu124 makes a nucleophilic attack on C1 atom of the transition state, forming a glycosyl-enzyme intermediate.
(5) Glu207 acts as a general base to activate a water molecule.
(6) The activated water makes a nucleophilic attack on the C1 atom of the intermediate. This reaction proceeds through an oxocarbenium-like transition state to complete the hydrolysis.

Created Updated
2004-05-17 2009-02-26