1bfn
From Proteopedia
(New page: 200px<br /><applet load="1bfn" size="450" color="white" frame="true" align="right" spinBox="true" caption="1bfn, resolution 2.07Å" /> '''BETA-AMYLASE/BETA-CY...) |
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| - | [[Image:1bfn.gif|left|200px]]<br /><applet load="1bfn" size=" | + | [[Image:1bfn.gif|left|200px]]<br /><applet load="1bfn" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1bfn, resolution 2.07Å" /> | caption="1bfn, resolution 2.07Å" /> | ||
'''BETA-AMYLASE/BETA-CYCLODEXTRIN COMPLEX'''<br /> | '''BETA-AMYLASE/BETA-CYCLODEXTRIN COMPLEX'''<br /> | ||
==Overview== | ==Overview== | ||
| - | In order to study the interaction of soybean beta-amylase with substrate, we solved the crystal structure of beta-cyclodextrin-enzyme complex and | + | In order to study the interaction of soybean beta-amylase with substrate, we solved the crystal structure of beta-cyclodextrin-enzyme complex and compared it with that of alpha-cyclodextrin-enzyme complex. The enzyme was expressed in Escherichia coli at a high level as a soluble and catalytically active protein. The purified recombinant enzyme had properties nearly identical to those of native soybean beta-amylase and formed the same crystals as the native enzyme. The crystal structure of recombinant enzyme complexed with beta-cyclodextrin was refined at 2. 07-A resolution with a final crystallographic R value of 15.8% (Rfree = 21.1%). The root mean square deviation in the position of C-alpha atoms between this recombinant enzyme and the native enzyme was 0.22 A. These results indicate that the expression system established here is suitable for studying structure-function relationships of beta-amylase. The conformation of the bound beta-cyclodextrin takes an ellipsoid shape in contrast to the circular shape of the bound alpha-cyclodextrin. The cyclodextrins shared mainly two glucose binding sites, 3 and 4. The glucose residue 4 was slightly shifted from the maltose binding site. This suggests that the binding site of the cyclodextrins is important for its holding of a cleaved substrate, which enables the multiple attack mechanism of beta-amylase. |
==About this Structure== | ==About this Structure== | ||
| - | 1BFN is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Glycine_max Glycine max] with SO4 as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Beta-amylase Beta-amylase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.2 3.2.1.2] Full crystallographic information is available from [http:// | + | 1BFN is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Glycine_max Glycine max] with <scene name='pdbligand=SO4:'>SO4</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Beta-amylase Beta-amylase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.2 3.2.1.2] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1BFN OCA]. |
==Reference== | ==Reference== | ||
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[[Category: recombinant]] | [[Category: recombinant]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 11:54:47 2008'' |
Revision as of 09:54, 21 February 2008
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BETA-AMYLASE/BETA-CYCLODEXTRIN COMPLEX
Overview
In order to study the interaction of soybean beta-amylase with substrate, we solved the crystal structure of beta-cyclodextrin-enzyme complex and compared it with that of alpha-cyclodextrin-enzyme complex. The enzyme was expressed in Escherichia coli at a high level as a soluble and catalytically active protein. The purified recombinant enzyme had properties nearly identical to those of native soybean beta-amylase and formed the same crystals as the native enzyme. The crystal structure of recombinant enzyme complexed with beta-cyclodextrin was refined at 2. 07-A resolution with a final crystallographic R value of 15.8% (Rfree = 21.1%). The root mean square deviation in the position of C-alpha atoms between this recombinant enzyme and the native enzyme was 0.22 A. These results indicate that the expression system established here is suitable for studying structure-function relationships of beta-amylase. The conformation of the bound beta-cyclodextrin takes an ellipsoid shape in contrast to the circular shape of the bound alpha-cyclodextrin. The cyclodextrins shared mainly two glucose binding sites, 3 and 4. The glucose residue 4 was slightly shifted from the maltose binding site. This suggests that the binding site of the cyclodextrins is important for its holding of a cleaved substrate, which enables the multiple attack mechanism of beta-amylase.
About this Structure
1BFN is a Single protein structure of sequence from Glycine max with as ligand. Active as Beta-amylase, with EC number 3.2.1.2 Full crystallographic information is available from OCA.
Reference
Crystal structure of recombinant soybean beta-amylase complexed with beta-cyclodextrin., Adachi M, Mikami B, Katsube T, Utsumi S, J Biol Chem. 1998 Jul 31;273(31):19859-65. PMID:9677422
Page seeded by OCA on Thu Feb 21 11:54:47 2008
Categories: Beta-amylase | Glycine max | Single protein | Adachi, M. | Katsube, T. | Mikami, B. | Utsumi, S. | SO4 | Beta-cyclodextrin | Hydrolase | Recombinant
