2e8z
From Proteopedia
(New page: 200px<br /><applet load="2e8z" size="350" color="white" frame="true" align="right" spinBox="true" caption="2e8z, resolution 2.20Å" /> '''Crystal structure of...) |
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| - | [[Image:2e8z.jpg|left|200px]] | + | [[Image:2e8z.jpg|left|200px]] |
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| - | '''Crystal structure of pullulanase type I from Bacillus subtilis str. 168 complexed with alpha-cyclodextrin''' | + | {{Structure |
| + | |PDB= 2e8z |SIZE=350|CAPTION= <scene name='initialview01'>2e8z</scene>, resolution 2.20Å | ||
| + | |SITE= <scene name='pdbsite=AC1:Glc+Binding+Site+For+Residue+A+721'>AC1</scene>, <scene name='pdbsite=AC2:Glc+Binding+Site+For+Residue+A+722'>AC2</scene>, <scene name='pdbsite=AC3:Glc+Binding+Site+For+Residue+A+723'>AC3</scene>, <scene name='pdbsite=AC4:Glc+Binding+Site+For+Residue+A+724'>AC4</scene>, <scene name='pdbsite=AC5:Glc+Binding+Site+For+Residue+A+725'>AC5</scene>, <scene name='pdbsite=AC6:Glc+Binding+Site+For+Residue+A+726'>AC6</scene>, <scene name='pdbsite=AC7:Glc+Binding+Site+For+Residue+B+731'>AC7</scene>, <scene name='pdbsite=AC8:Glc+Binding+Site+For+Residue+B+732'>AC8</scene>, <scene name='pdbsite=AC9:Glc+Binding+Site+For+Residue+B+733'>AC9</scene>, <scene name='pdbsite=BC1:Glc+Binding+Site+For+Residue+B+734'>BC1</scene>, <scene name='pdbsite=BC2:Glc+Binding+Site+For+Residue+B+735'>BC2</scene>, <scene name='pdbsite=BC3:Glc+Binding+Site+For+Residue+B+736'>BC3</scene>, <scene name='pdbsite=BC4:Ca+Binding+Site+For+Residue+A+741'>BC4</scene>, <scene name='pdbsite=BC5:Ca+Binding+Site+For+Residue+B+742'>BC5</scene>, <scene name='pdbsite=BC6:Act+Binding+Site+For+Residue+A+751'>BC6</scene>, <scene name='pdbsite=BC7:Act+Binding+Site+For+Residue+B+752'>BC7</scene>, <scene name='pdbsite=BC8:Gol+Binding+Site+For+Residue+A+761'>BC8</scene> and <scene name='pdbsite=BC9:Gol+Binding+Site+For+Residue+B+762'>BC9</scene> | ||
| + | |LIGAND= <scene name='pdbligand=CA:CALCIUM+ION'>CA</scene>, <scene name='pdbligand=ACT:ACETATE+ION'>ACT</scene> and <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene> | ||
| + | |ACTIVITY= [http://en.wikipedia.org/wiki/Pullulanase Pullulanase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.41 3.2.1.41] | ||
| + | |GENE= amyX ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=1423 Bacillus subtilis]) | ||
| + | }} | ||
| + | |||
| + | '''Crystal structure of pullulanase type I from Bacillus subtilis str. 168 complexed with alpha-cyclodextrin''' | ||
| + | |||
==Overview== | ==Overview== | ||
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==About this Structure== | ==About this Structure== | ||
| - | 2E8Z is a [ | + | 2E8Z is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Bacillus_subtilis Bacillus subtilis]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2E8Z OCA]. |
==Reference== | ==Reference== | ||
| - | Overexpression, purification and preliminary X-ray analysis of pullulanase from Bacillus subtilis strain 168., Malle D, Itoh T, Hashimoto W, Murata K, Utsumi S, Mikami B, Acta Crystallogr Sect F Struct Biol Cryst Commun. 2006 Apr 1;62(Pt, 4):381-4. Epub 2006 Mar 25. PMID:[http:// | + | Overexpression, purification and preliminary X-ray analysis of pullulanase from Bacillus subtilis strain 168., Malle D, Itoh T, Hashimoto W, Murata K, Utsumi S, Mikami B, Acta Crystallogr Sect F Struct Biol Cryst Commun. 2006 Apr 1;62(Pt, 4):381-4. Epub 2006 Mar 25. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/16582490 16582490] |
[[Category: Bacillus subtilis]] | [[Category: Bacillus subtilis]] | ||
[[Category: Pullulanase]] | [[Category: Pullulanase]] | ||
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[[Category: multiple domain]] | [[Category: multiple domain]] | ||
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Mar 20 16:37:00 2008'' |
Revision as of 14:37, 20 March 2008
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| , resolution 2.20Å | |||||||
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| Sites: | , , , , , , , , , , , , , , , , and | ||||||
| Ligands: | , and | ||||||
| Gene: | amyX (Bacillus subtilis) | ||||||
| Activity: | Pullulanase, with EC number 3.2.1.41 | ||||||
| Coordinates: | save as pdb, mmCIF, xml | ||||||
Crystal structure of pullulanase type I from Bacillus subtilis str. 168 complexed with alpha-cyclodextrin
Overview
The AmyX gene encoding pullulanase from the common spore-forming bacterium Bacillus subtilis strain 168 was cloned, overexpressed in Escherichia coli, purified and crystallized. The recombinant pullulanase was purified to homogeneity using ammonium sulfate precipitation, hydrophobic chromatography and anion-exchange chromatography, resulting in a specific activity of 24.10 U per milligram of protein. SDS-PAGE analysis showed that the molecular weight of the protein is approximately 81.0 kDa, which is similar to the calculated molecular weight, 81.1 kDa, from its translated cDNA sequence. The k(cat) and K(m) of the purified enzyme with pullulan as substrate were approximately 79 s(-1) and 1.284 mg ml(-1), respectively. X-ray crystallographic analysis of the pullulanase crystal showed that the crystal belongs to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 70.568, b = 127.68, c = 189.25 angstroms. The crystal contains two molecules of pullulanase in the asymmetric unit, with a solvent content of 53.15%. The crystal diffracted to 2.1 angstroms resolution at a synchrotron and is suitable for structure determination.
About this Structure
2E8Z is a Single protein structure of sequence from Bacillus subtilis. Full crystallographic information is available from OCA.
Reference
Overexpression, purification and preliminary X-ray analysis of pullulanase from Bacillus subtilis strain 168., Malle D, Itoh T, Hashimoto W, Murata K, Utsumi S, Mikami B, Acta Crystallogr Sect F Struct Biol Cryst Commun. 2006 Apr 1;62(Pt, 4):381-4. Epub 2006 Mar 25. PMID:16582490
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