1axk
From Proteopedia
(New page: 200px<br /><applet load="1axk" size="450" color="white" frame="true" align="right" spinBox="true" caption="1axk, resolution 2.1Å" /> '''ENGINEERED BACILLUS B...) |
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| - | [[Image:1axk.gif|left|200px]]<br /><applet load="1axk" size=" | + | [[Image:1axk.gif|left|200px]]<br /><applet load="1axk" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1axk, resolution 2.1Å" /> | caption="1axk, resolution 2.1Å" /> | ||
'''ENGINEERED BACILLUS BIFUNCTIONAL ENZYME GLUXYN-1'''<br /> | '''ENGINEERED BACILLUS BIFUNCTIONAL ENZYME GLUXYN-1'''<br /> | ||
==Overview== | ==Overview== | ||
| - | The 1,3-1,4-beta-glucanase from Bacillus macerans (wtGLU) and the 1, 4-beta-xylanase from Bacillus subtilis (wtXYN) are both single-domain | + | The 1,3-1,4-beta-glucanase from Bacillus macerans (wtGLU) and the 1, 4-beta-xylanase from Bacillus subtilis (wtXYN) are both single-domain jellyroll proteins catalyzing similar enzymatic reactions. In the fusion protein GluXyn-1, the two proteins are joined by insertion of the entire XYN domain into a surface loop of cpMAC-57, a circularly permuted variant of wtGLU. GluXyn-1 was generated by protein engineering methods, produced in Escherichia coli and shown to fold spontaneously and have both enzymatic activities at wild-type level. The crystal structure of GluXyn-1 was determined at 2.1 A resolution and refined to R = 17.7% and R(free) = 22.4%. It shows nearly ideal, native-like folding of both protein domains and a small, but significant hinge bending between the domains. The active sites are independent and accessible explaining the observed enzymatic activity. Because in GluXyn-1 the complete XYN domain is inserted into the compact folding unit of GLU, the wild-type-like activity and tertiary structure of the latter proves that the folding process of GLU does not depend on intramolecular interactions that are short-ranged in the sequence. Insertion fusions of the GluXyn-1 type may prove to be an easy route toward more stable bifunctional proteins in which the two parts are more closely associated than in linear end-to-end protein fusions. |
==About this Structure== | ==About this Structure== | ||
| - | 1AXK is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Bacillus_subtilis Bacillus subtilis] with CA as [http://en.wikipedia.org/wiki/ligand ligand]. Full crystallographic information is available from [http:// | + | 1AXK is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Bacillus_subtilis Bacillus subtilis] with <scene name='pdbligand=CA:'>CA</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1AXK OCA]. |
==Reference== | ==Reference== | ||
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[[Category: hybrid enzyme]] | [[Category: hybrid enzyme]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 11:49:20 2008'' |
Revision as of 09:49, 21 February 2008
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ENGINEERED BACILLUS BIFUNCTIONAL ENZYME GLUXYN-1
Overview
The 1,3-1,4-beta-glucanase from Bacillus macerans (wtGLU) and the 1, 4-beta-xylanase from Bacillus subtilis (wtXYN) are both single-domain jellyroll proteins catalyzing similar enzymatic reactions. In the fusion protein GluXyn-1, the two proteins are joined by insertion of the entire XYN domain into a surface loop of cpMAC-57, a circularly permuted variant of wtGLU. GluXyn-1 was generated by protein engineering methods, produced in Escherichia coli and shown to fold spontaneously and have both enzymatic activities at wild-type level. The crystal structure of GluXyn-1 was determined at 2.1 A resolution and refined to R = 17.7% and R(free) = 22.4%. It shows nearly ideal, native-like folding of both protein domains and a small, but significant hinge bending between the domains. The active sites are independent and accessible explaining the observed enzymatic activity. Because in GluXyn-1 the complete XYN domain is inserted into the compact folding unit of GLU, the wild-type-like activity and tertiary structure of the latter proves that the folding process of GLU does not depend on intramolecular interactions that are short-ranged in the sequence. Insertion fusions of the GluXyn-1 type may prove to be an easy route toward more stable bifunctional proteins in which the two parts are more closely associated than in linear end-to-end protein fusions.
About this Structure
1AXK is a Single protein structure of sequence from Bacillus subtilis with as ligand. Full crystallographic information is available from OCA.
Reference
Structure and function of the Bacillus hybrid enzyme GluXyn-1: native-like jellyroll fold preserved after insertion of autonomous globular domain., Ay J, Gotz F, Borriss R, Heinemann U, Proc Natl Acad Sci U S A. 1998 Jun 9;95(12):6613-8. PMID:9618460
Page seeded by OCA on Thu Feb 21 11:49:20 2008
Categories: Bacillus subtilis | Single protein | Ay, J. | Heinemann, U. | CA | 1 | 3-1 | 4-beta-glucanase | 4-beta-xylanase | Bifunctional | Fusion protein | Gluxyn-1 | Hybrid enzyme
