![[Main Page]](/wiki/logos/proteopedia_135x200px_v1.01_15Years.png)
(New page: 200px<br /><applet load="1cxi" size="450" color="white" frame="true" align="right" spinBox="true" caption="1cxi, resolution 2.2Å" /> '''WILD-TYPE CGTASE FROM...) |
|||
| Line 1: | Line 1: | ||
| - | [[Image:1cxi.gif|left|200px]]<br /><applet load="1cxi" size=" | + | [[Image:1cxi.gif|left|200px]]<br /><applet load="1cxi" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1cxi, resolution 2.2Å" /> | caption="1cxi, resolution 2.2Å" /> | ||
'''WILD-TYPE CGTASE FROM BACILLUS CIRCULANS STRAIN 251 AT 120 K AND PH 7.55'''<br /> | '''WILD-TYPE CGTASE FROM BACILLUS CIRCULANS STRAIN 251 AT 120 K AND PH 7.55'''<br /> | ||
==Overview== | ==Overview== | ||
| - | Asp-229, Glu-257, and Asp-328 constitute the catalytic residues in | + | Asp-229, Glu-257, and Asp-328 constitute the catalytic residues in cyclodextrin glycosyl transferase from Bacillus circulans strain 251. Via site-directed mutagenesis constructed D229N, E257Q, and D328N mutant proteins showed a 4,000-60,000-fold reduction of cyclization activity. A D229N/E257Q double mutant showed a 700,000-fold reduction and was crystallized for use in soaking experiments with alpha-cyclodextrin. Crystal structures were determined of wild type CGTase soaked at elevated pH with alpha-cyclodextrin (resolution, 2.1 A) and maltoheptaose (2.4 A). In addition, structures at cryogenic temperature were solved of the unliganded enzyme (2.2 A) and of the D229N/E257Q mutant after soaking with alpha-cyclodextrin (2.6 A). In the crystals soaked in alpha-cyclodextrin and maltoheptaose, a maltotetraose molecule is observed to bind in the active site. Residue 229 is at hydrogen bonding distance from the C-6 hydroxyl group of the sugar, which after cleavage will contain the new reducing end. In the D229N/E257Q double mutant structure, two alpha-cyclodextrins are observed to replace two maltoses at the E-domain, thus providing structural information on product inhibition via binding to the enzyme's raw starch binding domain. |
==About this Structure== | ==About this Structure== | ||
| - | 1CXI is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Bacillus_circulans Bacillus circulans] with MAL and CA as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Cyclomaltodextrin_glucanotransferase Cyclomaltodextrin glucanotransferase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.4.1.19 2.4.1.19] Full crystallographic information is available from [http:// | + | 1CXI is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Bacillus_circulans Bacillus circulans] with <scene name='pdbligand=MAL:'>MAL</scene> and <scene name='pdbligand=CA:'>CA</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Cyclomaltodextrin_glucanotransferase Cyclomaltodextrin glucanotransferase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.4.1.19 2.4.1.19] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1CXI OCA]. |
==Reference== | ==Reference== | ||
| Line 14: | Line 14: | ||
[[Category: Cyclomaltodextrin glucanotransferase]] | [[Category: Cyclomaltodextrin glucanotransferase]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
| - | [[Category: Dijkstra, B | + | [[Category: Dijkstra, B W.]] |
| - | [[Category: Knegtel, R | + | [[Category: Knegtel, R M.A.]] |
| - | [[Category: Strokopytov, B | + | [[Category: Strokopytov, B V.]] |
[[Category: CA]] | [[Category: CA]] | ||
[[Category: MAL]] | [[Category: MAL]] | ||
[[Category: glycosyltransferase]] | [[Category: glycosyltransferase]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 12:10:46 2008'' |
Revision as of 10:10, 21 February 2008
|
WILD-TYPE CGTASE FROM BACILLUS CIRCULANS STRAIN 251 AT 120 K AND PH 7.55
Overview
Asp-229, Glu-257, and Asp-328 constitute the catalytic residues in cyclodextrin glycosyl transferase from Bacillus circulans strain 251. Via site-directed mutagenesis constructed D229N, E257Q, and D328N mutant proteins showed a 4,000-60,000-fold reduction of cyclization activity. A D229N/E257Q double mutant showed a 700,000-fold reduction and was crystallized for use in soaking experiments with alpha-cyclodextrin. Crystal structures were determined of wild type CGTase soaked at elevated pH with alpha-cyclodextrin (resolution, 2.1 A) and maltoheptaose (2.4 A). In addition, structures at cryogenic temperature were solved of the unliganded enzyme (2.2 A) and of the D229N/E257Q mutant after soaking with alpha-cyclodextrin (2.6 A). In the crystals soaked in alpha-cyclodextrin and maltoheptaose, a maltotetraose molecule is observed to bind in the active site. Residue 229 is at hydrogen bonding distance from the C-6 hydroxyl group of the sugar, which after cleavage will contain the new reducing end. In the D229N/E257Q double mutant structure, two alpha-cyclodextrins are observed to replace two maltoses at the E-domain, thus providing structural information on product inhibition via binding to the enzyme's raw starch binding domain.
About this Structure
1CXI is a Single protein structure of sequence from Bacillus circulans with and as ligands. Active as Cyclomaltodextrin glucanotransferase, with EC number 2.4.1.19 Full crystallographic information is available from OCA.
Reference
Crystallographic studies of the interaction of cyclodextrin glycosyltransferase from Bacillus circulans strain 251 with natural substrates and products., Knegtel RM, Strokopytov B, Penninga D, Faber OG, Rozeboom HJ, Kalk KH, Dijkhuizen L, Dijkstra BW, J Biol Chem. 1995 Dec 8;270(49):29256-64. PMID:7493956
Page seeded by OCA on Thu Feb 21 12:10:46 2008