1mw3
From Proteopedia
(New page: 200px<br /><applet load="1mw3" size="450" color="white" frame="true" align="right" spinBox="true" caption="1mw3, resolution 2.00Å" /> '''Amylosucrase soaked ...) |
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| - | [[Image:1mw3.jpg|left|200px]]<br /><applet load="1mw3" size=" | + | [[Image:1mw3.jpg|left|200px]]<br /><applet load="1mw3" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1mw3, resolution 2.00Å" /> | caption="1mw3, resolution 2.00Å" /> | ||
'''Amylosucrase soaked with 1M sucrose'''<br /> | '''Amylosucrase soaked with 1M sucrose'''<br /> | ||
==Overview== | ==Overview== | ||
| - | The glucosyltransferase amylosucrase is structurally quite similar to the | + | The glucosyltransferase amylosucrase is structurally quite similar to the hydrolase alpha-amylase. How this switch in functionality is achieved is an important and fundamental question. The inactive E328Q amylosucrase variant has been co-crystallized with maltoheptaose, and the structure was determined by x-ray crystallography to 2.2 A resolution, revealing a maltoheptaose binding site in the B'-domain somewhat distant from the active site. Additional soaking of these crystals with maltoheptaose resulted in replacement of Tris in the active site with maltoheptaose, allowing the mapping of the -1 to +5 binding subsites. Crystals of amylosucrase were soaked with sucrose at different concentrations. The structures at approximately 2.1 A resolution revealed three new binding sites of different affinity. The highest affinity binding site is close to the active site but is not in the previously identified substrate access channel. Allosteric regulation seems necessary to facilitate access from this binding site. The structures show the pivotal role of the B'-domain in the transferase reaction. Based on these observations, an extension of the hydrolase reaction mechanism valid for this enzyme can be proposed. In this mechanism, the glycogen-like polymer is bound in the widest access channel to the active site. The polymer binding introduces structural changes that allow sucrose to migrate from its binding site into the active site and displace the polymer. |
==About this Structure== | ==About this Structure== | ||
| - | 1MW3 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Neisseria_polysaccharea Neisseria polysaccharea] with SUC and TRS as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Amylosucrase Amylosucrase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.4.1.4 2.4.1.4] Full crystallographic information is available from [http:// | + | 1MW3 is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Neisseria_polysaccharea Neisseria polysaccharea] with <scene name='pdbligand=SUC:'>SUC</scene> and <scene name='pdbligand=TRS:'>TRS</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Amylosucrase Amylosucrase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.4.1.4 2.4.1.4] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1MW3 OCA]. |
==Reference== | ==Reference== | ||
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[[Category: Monsan, P.]] | [[Category: Monsan, P.]] | ||
[[Category: Remaud-Simeon, M.]] | [[Category: Remaud-Simeon, M.]] | ||
| - | [[Category: Skov, L | + | [[Category: Skov, L K.]] |
[[Category: Sprogoe, D.]] | [[Category: Sprogoe, D.]] | ||
[[Category: SUC]] | [[Category: SUC]] | ||
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[[Category: protein-sugar complex]] | [[Category: protein-sugar complex]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 13:59:47 2008'' |
Revision as of 11:59, 21 February 2008
|
Amylosucrase soaked with 1M sucrose
Overview
The glucosyltransferase amylosucrase is structurally quite similar to the hydrolase alpha-amylase. How this switch in functionality is achieved is an important and fundamental question. The inactive E328Q amylosucrase variant has been co-crystallized with maltoheptaose, and the structure was determined by x-ray crystallography to 2.2 A resolution, revealing a maltoheptaose binding site in the B'-domain somewhat distant from the active site. Additional soaking of these crystals with maltoheptaose resulted in replacement of Tris in the active site with maltoheptaose, allowing the mapping of the -1 to +5 binding subsites. Crystals of amylosucrase were soaked with sucrose at different concentrations. The structures at approximately 2.1 A resolution revealed three new binding sites of different affinity. The highest affinity binding site is close to the active site but is not in the previously identified substrate access channel. Allosteric regulation seems necessary to facilitate access from this binding site. The structures show the pivotal role of the B'-domain in the transferase reaction. Based on these observations, an extension of the hydrolase reaction mechanism valid for this enzyme can be proposed. In this mechanism, the glycogen-like polymer is bound in the widest access channel to the active site. The polymer binding introduces structural changes that allow sucrose to migrate from its binding site into the active site and displace the polymer.
About this Structure
1MW3 is a Single protein structure of sequence from Neisseria polysaccharea with and as ligands. Active as Amylosucrase, with EC number 2.4.1.4 Full crystallographic information is available from OCA.
Reference
Oligosaccharide and sucrose complexes of amylosucrase. Structural implications for the polymerase activity., Skov LK, Mirza O, Sprogoe D, Dar I, Remaud-Simeon M, Albenne C, Monsan P, Gajhede M, J Biol Chem. 2002 Dec 6;277(49):47741-7. Epub 2002 Oct 2. PMID:12364331
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