1xwt
From Proteopedia
(New page: 200px<br /><applet load="1xwt" size="450" color="white" frame="true" align="right" spinBox="true" caption="1xwt, resolution 1.30Å" /> '''Structure Of A Cold-...) |
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| - | [[Image:1xwt.gif|left|200px]]<br /><applet load="1xwt" size=" | + | [[Image:1xwt.gif|left|200px]]<br /><applet load="1xwt" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1xwt, resolution 1.30Å" /> | caption="1xwt, resolution 1.30Å" /> | ||
'''Structure Of A Cold-Adapted Family 8 Xylanase'''<br /> | '''Structure Of A Cold-Adapted Family 8 Xylanase'''<br /> | ||
==Overview== | ==Overview== | ||
| - | Site-directed mutagenesis and a comparative characterisation of the | + | Site-directed mutagenesis and a comparative characterisation of the kinetic parameters, pH dependency of activity and thermal stability of mutant and wild-type enzymes have been used in association with crystallographic analysis to delineate the functions of several active site residues in a novel glycoside hydrolase family 8 xylanase. Each of the residues investigated plays an essential role in this enzyme: E78 as the general acid, D281 as the general base and in orientating the nucleophilic water molecule, Y203 in maintaining the position of the nucleophilic water molecule and in structural integrity and D144 in sugar ring distortion and transition state stabilization. Interestingly, although crystal structure analyses and the pH-activity profiles clearly identify the functions of E78 and D281, substitution of these residues with their amide derivatives results in only a 250-fold and 700-fold reduction in their apparent k(cat) values, respectively. This, in addition to the observation that the proposed general base is not conserved in all glycoside hydrolase family 8 enzymes, indicates that the mechanistic architecture in this family of inverting enzymes is more complex than is conventionally believed and points to a diversity in the identity of the mechanistically important residues as well as in the arrangement of the intricate microenvironment of the active site among members of this family. |
==About this Structure== | ==About this Structure== | ||
| - | 1XWT is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Pseudoalteromonas_haloplanktis Pseudoalteromonas haloplanktis]. Active as [http://en.wikipedia.org/wiki/Endo-1,4-beta-xylanase Endo-1,4-beta-xylanase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.8 3.2.1.8] Full crystallographic information is available from [http:// | + | 1XWT is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Pseudoalteromonas_haloplanktis Pseudoalteromonas haloplanktis]. Active as [http://en.wikipedia.org/wiki/Endo-1,4-beta-xylanase Endo-1,4-beta-xylanase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.8 3.2.1.8] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1XWT OCA]. |
==Reference== | ==Reference== | ||
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[[Category: Pseudoalteromonas haloplanktis]] | [[Category: Pseudoalteromonas haloplanktis]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
| - | [[Category: Beeumen, J | + | [[Category: Beeumen, J J.Van.]] |
[[Category: Collins, T.]] | [[Category: Collins, T.]] | ||
[[Category: Feller, G.]] | [[Category: Feller, G.]] | ||
[[Category: Gerday, C.]] | [[Category: Gerday, C.]] | ||
[[Category: Hoyoux, A.]] | [[Category: Hoyoux, A.]] | ||
| - | [[Category: Savvides, S | + | [[Category: Savvides, S N.]] |
| - | [[Category: Vos, D | + | [[Category: Vos, D De.]] |
[[Category: cold adaptation]] | [[Category: cold adaptation]] | ||
[[Category: family 8]] | [[Category: family 8]] | ||
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[[Category: xylan degradation]] | [[Category: xylan degradation]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 15:59:26 2008'' |
Revision as of 13:59, 21 February 2008
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Structure Of A Cold-Adapted Family 8 Xylanase
Overview
Site-directed mutagenesis and a comparative characterisation of the kinetic parameters, pH dependency of activity and thermal stability of mutant and wild-type enzymes have been used in association with crystallographic analysis to delineate the functions of several active site residues in a novel glycoside hydrolase family 8 xylanase. Each of the residues investigated plays an essential role in this enzyme: E78 as the general acid, D281 as the general base and in orientating the nucleophilic water molecule, Y203 in maintaining the position of the nucleophilic water molecule and in structural integrity and D144 in sugar ring distortion and transition state stabilization. Interestingly, although crystal structure analyses and the pH-activity profiles clearly identify the functions of E78 and D281, substitution of these residues with their amide derivatives results in only a 250-fold and 700-fold reduction in their apparent k(cat) values, respectively. This, in addition to the observation that the proposed general base is not conserved in all glycoside hydrolase family 8 enzymes, indicates that the mechanistic architecture in this family of inverting enzymes is more complex than is conventionally believed and points to a diversity in the identity of the mechanistically important residues as well as in the arrangement of the intricate microenvironment of the active site among members of this family.
About this Structure
1XWT is a Single protein structure of sequence from Pseudoalteromonas haloplanktis. Active as Endo-1,4-beta-xylanase, with EC number 3.2.1.8 Full crystallographic information is available from OCA.
Reference
Study of the active site residues of a glycoside hydrolase family 8 xylanase., Collins T, De Vos D, Hoyoux A, Savvides SN, Gerday C, Van Beeumen J, Feller G, J Mol Biol. 2005 Nov 25;354(2):425-35. Epub 2005 Oct 10. PMID:16246370
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