1x0c
From Proteopedia
(New page: 200px<br /><applet load="1x0c" size="450" color="white" frame="true" align="right" spinBox="true" caption="1x0c, resolution 1.70Å" /> '''Improved Crystal Str...) |
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| - | [[Image:1x0c.gif|left|200px]]<br /><applet load="1x0c" size=" | + | [[Image:1x0c.gif|left|200px]]<br /><applet load="1x0c" size="350" color="white" frame="true" align="right" spinBox="true" |
caption="1x0c, resolution 1.70Å" /> | caption="1x0c, resolution 1.70Å" /> | ||
'''Improved Crystal Structure of Isopullulanase from Aspergillus niger ATCC 9642'''<br /> | '''Improved Crystal Structure of Isopullulanase from Aspergillus niger ATCC 9642'''<br /> | ||
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| + | ==Overview== | ||
| + | An isopullulanase (IPU) from Aspergillus niger ATCC9642 hydrolyzes, alpha-1,4-glucosidic linkages of pullulan to produce isopanose. Although, IPU does not hydrolyze dextran, it is classified into glycoside hydrolase, family 49 (GH49), major members of which are dextran-hydrolyzing enzymes., IPU is highly glycosylated, making it difficult to obtain its crystal. We, used endoglycosidase H(f) to cleave the N-linked oligosaccharides of IPU, and we here determined the unliganded and isopanose-complexed forms of, IPU, both solved at 1.7-A resolution. IPU is composed of domains N and C, joined by a short linker, with electron density maps for 11 or 12, N-acetylglucosamine residues per molecule. Domain N consists of 13, beta-strands and forms a beta-sandwich. Domain C, where the active site is, located, forms a right-handed beta-helix, and the lengths of the pitches, of each coil of the beta-helix are similar to those of GH49 dextranase and, GH28 polygalacturonase. The entire structure of IPU resembles that of a, GH49 enzyme, Penicillium minioluteum dextranase (Dex49A), despite a, difference in substrate specificity. Compared with the active sites of IPU, and Dex49A, the amino acid residues participating in subsites +2 and +3, are not conserved, and the glucose residues of isopanose bound to IPU, completely differ in orientation from the corresponding glucose residues, of isomaltose bound to Dex49A. The shape of the catalytic cleft, characterized by the seventh coil of the beta-helix and a loop from domain, N appears to be critical in determining the specificity of IPU for, pullulan. | ||
==About this Structure== | ==About this Structure== | ||
| - | 1X0C is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Aspergillus_niger Aspergillus niger] with NAG as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Isopullulanase Isopullulanase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.57 3.2.1.57] Full crystallographic information is available from [http:// | + | 1X0C is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Aspergillus_niger Aspergillus niger] with <scene name='pdbligand=NAG:'>NAG</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. Active as [http://en.wikipedia.org/wiki/Isopullulanase Isopullulanase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.2.1.57 3.2.1.57] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1X0C OCA]. |
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| + | ==Reference== | ||
| + | Crystal structure of Aspergillus niger isopullulanase, a member of glycoside hydrolase family 49., Mizuno M, Koide A, Yamamura A, Akeboshi H, Yoshida H, Kamitori S, Sakano Y, Nishikawa A, Tonozuka T, J Mol Biol. 2008 Feb 8;376(1):210-20. Epub 2007 Dec 5. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=18155243 18155243] | ||
[[Category: Aspergillus niger]] | [[Category: Aspergillus niger]] | ||
[[Category: Isopullulanase]] | [[Category: Isopullulanase]] | ||
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[[Category: pullulan]] | [[Category: pullulan]] | ||
| - | ''Page seeded by [http:// | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Wed Jan 23 10:36:07 2008'' |
Revision as of 08:36, 23 January 2008
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Improved Crystal Structure of Isopullulanase from Aspergillus niger ATCC 9642
Overview
An isopullulanase (IPU) from Aspergillus niger ATCC9642 hydrolyzes, alpha-1,4-glucosidic linkages of pullulan to produce isopanose. Although, IPU does not hydrolyze dextran, it is classified into glycoside hydrolase, family 49 (GH49), major members of which are dextran-hydrolyzing enzymes., IPU is highly glycosylated, making it difficult to obtain its crystal. We, used endoglycosidase H(f) to cleave the N-linked oligosaccharides of IPU, and we here determined the unliganded and isopanose-complexed forms of, IPU, both solved at 1.7-A resolution. IPU is composed of domains N and C, joined by a short linker, with electron density maps for 11 or 12, N-acetylglucosamine residues per molecule. Domain N consists of 13, beta-strands and forms a beta-sandwich. Domain C, where the active site is, located, forms a right-handed beta-helix, and the lengths of the pitches, of each coil of the beta-helix are similar to those of GH49 dextranase and, GH28 polygalacturonase. The entire structure of IPU resembles that of a, GH49 enzyme, Penicillium minioluteum dextranase (Dex49A), despite a, difference in substrate specificity. Compared with the active sites of IPU, and Dex49A, the amino acid residues participating in subsites +2 and +3, are not conserved, and the glucose residues of isopanose bound to IPU, completely differ in orientation from the corresponding glucose residues, of isomaltose bound to Dex49A. The shape of the catalytic cleft, characterized by the seventh coil of the beta-helix and a loop from domain, N appears to be critical in determining the specificity of IPU for, pullulan.
About this Structure
1X0C is a Single protein structure of sequence from Aspergillus niger with as ligand. Active as Isopullulanase, with EC number 3.2.1.57 Full crystallographic information is available from OCA.
Reference
Crystal structure of Aspergillus niger isopullulanase, a member of glycoside hydrolase family 49., Mizuno M, Koide A, Yamamura A, Akeboshi H, Yoshida H, Kamitori S, Sakano Y, Nishikawa A, Tonozuka T, J Mol Biol. 2008 Feb 8;376(1):210-20. Epub 2007 Dec 5. PMID:18155243
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