1qi2: Difference between revisions

Line 1:

[[Image:1qi2.jpg|left|200px]]<br /><applet load="1qi2" size="~~450~~" color="white" frame="true" align="right" spinBox="true"

[[Image:1qi2.jpg|left|200px]]<br /><applet load="1qi2" size="350" color="white" frame="true" align="right" spinBox="true"

caption="1qi2, resolution 1.750Å" />

'''ENDOGLUCANASE CEL5A FROM BACILLUS AGARADHAERENS IN THE TETRAGONAL CRYSTAL FORM IN COMPLEX WITH 2',4'-DINITROPHENYL 2-DEOXY-2-FLUORO-B-D-CELLOTRIOSIDE'''<br />

==Overview==

Glycoside hydrolases are ubiquitous enzymes involved in a diverse array of, biological processes, from the breakdown of biomass, through to viral, invasion and cellular signalling. Endoglucanase Cel5A from Bacillus, agaradhaerens, classified into glycoside hydrolase family 5, has been, studied in a catalytically inactive crystal form at low pH conditions, in, which native and complex structures revealed the importance of ring, distortion during catalysis. Here, we present the structure of Cel5A in a, new crystal form obtained at higher pH values in which the enzyme is, active "in-crystal". Native, cellotriosyl-enzyme intermediate and, beta-d-cellobiose structures were solved at 1.95, 1.75 and 2.1 A, resolution, respectively. These structures reveal two classes of, conformational change: those caused by crystal-packing and pH, with others, induced upon substrate binding. At pH 7 a histidine residue, His206, implicated in substrate-binding and catalysis, but previously far removed, from the substrate-binding cleft, moves over 10 A into the active site, cleft in order to interact with the substrate in the +2 subsite., Occupation of the -1 subsite by substrate induces a loop closure to, optimise protein-ligand interactions. Cel5A, along with the unrelated, family 45 and family 6 cellulases, provides further evidence of, substantial conformational change in response to ligand binding for this, class of hydrolytic enzyme.

Glycoside hydrolases are ubiquitous enzymes involved in a diverse array of biological processes, from the breakdown of biomass, through to viral invasion and cellular signalling. Endoglucanase Cel5A from Bacillus agaradhaerens, classified into glycoside hydrolase family 5, has been studied in a catalytically inactive crystal form at low pH conditions, in which native and complex structures revealed the importance of ring distortion during catalysis. Here, we present the structure of Cel5A in a new crystal form obtained at higher pH values in which the enzyme is active "in-crystal". Native, cellotriosyl-enzyme intermediate and beta-d-cellobiose structures were solved at 1.95, 1.75 and 2.1 A resolution, respectively. These structures reveal two classes of conformational change: those caused by crystal-packing and pH, with others induced upon substrate binding. At pH 7 a histidine residue, His206, implicated in substrate-binding and catalysis, but previously far removed from the substrate-binding cleft, moves over 10 A into the active site cleft in order to interact with the substrate in the +2 subsite. Occupation of the -1 subsite by substrate induces a loop closure to optimise protein-ligand interactions. Cel5A, along with the unrelated family 45 and family 6 cellulases, provides further evidence of substantial conformational change in response to ligand binding for this class of hydrolytic enzyme.

==About this Structure==

1QI2 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Bacillus_agaradhaerens Bacillus agaradhaerens] with CA and FCT as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Cellulase Cellulase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.4 3.2.1.4] Full crystallographic information is available from [http://~~ispc~~.weizmann.ac.il/oca-bin/ocashort?id=1QI2 OCA].

1QI2 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Bacillus_agaradhaerens Bacillus agaradhaerens] with <scene name='pdbligand=CA:'>CA</scene> and <scene name='pdbligand=FCT:'>FCT</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Cellulase Cellulase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.4 3.2.1.4] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1QI2 OCA].

==Reference==

Line 14:

[[Category: Cellulase]]

[[Category: Single protein]]

[[Category: Davies, G.J.]]

[[Category: Davies, G J.]]

[[Category: Schulein, M.]]

[[Category: Varrot, A.]]

Line 24:

[[Category: hydrolase]]

''Page seeded by [http://~~ispc~~.weizmann.ac.il/oca OCA ] on ~~Wed Nov~~ 21 00:43:~~43 2007~~''

''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 14:39:52 2008''

Navigation menu

Page actions

Page actions

Personal tools

Navigation

Search

Tools

@@ Line 1: / Line 1: @@
-[[Image:1qi2.jpg|left|200px]]<br /><applet load="1qi2" size="450" color="white" frame="true" align="right" spinBox="true"
+[[Image:1qi2.jpg|left|200px]]<br /><applet load="1qi2" size="350" color="white" frame="true" align="right" spinBox="true"
 caption="1qi2, resolution 1.750&Aring;" />
 '''ENDOGLUCANASE CEL5A FROM BACILLUS AGARADHAERENS IN THE TETRAGONAL CRYSTAL FORM IN COMPLEX WITH 2',4'-DINITROPHENYL 2-DEOXY-2-FLUORO-B-D-CELLOTRIOSIDE'''<br />
 ==Overview==
-Glycoside hydrolases are ubiquitous enzymes involved in a diverse array of, biological processes, from the breakdown of biomass, through to viral, invasion and cellular signalling. Endoglucanase Cel5A from Bacillus, agaradhaerens, classified into glycoside hydrolase family 5, has been, studied in a catalytically inactive crystal form at low pH conditions, in, which native and complex structures revealed the importance of ring, distortion during catalysis. Here, we present the structure of Cel5A in a, new crystal form obtained at higher pH values in which the enzyme is, active "in-crystal". Native, cellotriosyl-enzyme intermediate and, beta-d-cellobiose structures were solved at 1.95, 1.75 and 2.1 A, resolution, respectively. These structures reveal two classes of, conformational change: those caused by crystal-packing and pH, with others, induced upon substrate binding. At pH 7 a histidine residue, His206, implicated in substrate-binding and catalysis, but previously far removed, from the substrate-binding cleft, moves over 10 A into the active site, cleft in order to interact with the substrate in the +2 subsite., Occupation of the -1 subsite by substrate induces a loop closure to, optimise protein-ligand interactions. Cel5A, along with the unrelated, family 45 and family 6 cellulases, provides further evidence of, substantial conformational change in response to ligand binding for this, class of hydrolytic enzyme.
+Glycoside hydrolases are ubiquitous enzymes involved in a diverse array of biological processes, from the breakdown of biomass, through to viral invasion and cellular signalling. Endoglucanase Cel5A from Bacillus agaradhaerens, classified into glycoside hydrolase family 5, has been studied in a catalytically inactive crystal form at low pH conditions, in which native and complex structures revealed the importance of ring distortion during catalysis. Here, we present the structure of Cel5A in a new crystal form obtained at higher pH values in which the enzyme is active "in-crystal". Native, cellotriosyl-enzyme intermediate and beta-d-cellobiose structures were solved at 1.95, 1.75 and 2.1 A resolution, respectively. These structures reveal two classes of conformational change: those caused by crystal-packing and pH, with others induced upon substrate binding. At pH 7 a histidine residue, His206, implicated in substrate-binding and catalysis, but previously far removed from the substrate-binding cleft, moves over 10 A into the active site cleft in order to interact with the substrate in the +2 subsite. Occupation of the -1 subsite by substrate induces a loop closure to optimise protein-ligand interactions. Cel5A, along with the unrelated family 45 and family 6 cellulases, provides further evidence of substantial conformational change in response to ligand binding for this class of hydrolytic enzyme.
 ==About this Structure==
-QI2 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Bacillus_agaradhaerens Bacillus agaradhaerens] with CA and FCT as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Cellulase Cellulase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.4 3.2.1.4] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1QI2 OCA].
+QI2 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Bacillus_agaradhaerens Bacillus agaradhaerens] with <scene name='pdbligand=CA:'>CA</scene> and <scene name='pdbligand=FCT:'>FCT</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Cellulase Cellulase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.4 3.2.1.4] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1QI2 OCA].
 ==Reference==
@@ Line 14: / Line 14: @@
 [[Category: Cellulase]]
 [[Category: Single protein]]
-[[Category: Davies, G.J.]]
+[[Category: Davies, G J.]]
 [[Category: Schulein, M.]]
 [[Category: Varrot, A.]]
@@ Line 24: / Line 24: @@
 [[Category: hydrolase]]
-''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Wed Nov 21 00:43:43 2007''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 14:39:52 2008''