2ggl
From Proteopedia
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| - | [[Image:2ggl.gif|left|200px]] | + | [[Image:2ggl.gif|left|200px]] |
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| - | '''The mutant A222C of Agrobacterium radiobacter N-carbamoyl-D-amino acid amidohydrolase''' | + | {{Structure |
| + | |PDB= 2ggl |SIZE=350|CAPTION= <scene name='initialview01'>2ggl</scene>, resolution 2.40Å | ||
| + | |SITE= | ||
| + | |LIGAND= | ||
| + | |ACTIVITY= [http://en.wikipedia.org/wiki/N-carbamoyl-D-amino_acid_hydrolase N-carbamoyl-D-amino acid hydrolase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.5.1.77 3.5.1.77] | ||
| + | |GENE= | ||
| + | }} | ||
| + | |||
| + | '''The mutant A222C of Agrobacterium radiobacter N-carbamoyl-D-amino acid amidohydrolase''' | ||
| + | |||
==Overview== | ==Overview== | ||
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==About this Structure== | ==About this Structure== | ||
| - | 2GGL is a [ | + | 2GGL is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Agrobacterium_tumefaciens Agrobacterium tumefaciens]. This structure supersedes the now removed PDB entries 2FKU and 2BA4. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2GGL OCA]. |
==Reference== | ==Reference== | ||
| - | Structure-stability-activity relationship in covalently cross-linked N-carbamoyl D-amino acid amidohydrolase and N-acylamino acid racemase., Chiu WC, You JY, Liu JS, Hsu SK, Hsu WH, Shih CH, Hwang JK, Wang WC, J Mol Biol. 2006 Jun 9;359(3):741-53. Epub 2006 Apr 18. PMID:[http:// | + | Structure-stability-activity relationship in covalently cross-linked N-carbamoyl D-amino acid amidohydrolase and N-acylamino acid racemase., Chiu WC, You JY, Liu JS, Hsu SK, Hsu WH, Shih CH, Hwang JK, Wang WC, J Mol Biol. 2006 Jun 9;359(3):741-53. Epub 2006 Apr 18. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/16650857 16650857] |
[[Category: Agrobacterium tumefaciens]] | [[Category: Agrobacterium tumefaciens]] | ||
[[Category: N-carbamoyl-D-amino acid hydrolase]] | [[Category: N-carbamoyl-D-amino acid hydrolase]] | ||
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[[Category: n-carbamoyl-d-amino-acid amidohydrolase]] | [[Category: n-carbamoyl-d-amino-acid amidohydrolase]] | ||
| - | ''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 17:04:31 2008'' |
Revision as of 15:04, 20 March 2008
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| , resolution 2.40Å | |||||||
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| Activity: | N-carbamoyl-D-amino acid hydrolase, with EC number 3.5.1.77 | ||||||
| Coordinates: | save as pdb, mmCIF, xml | ||||||
The mutant A222C of Agrobacterium radiobacter N-carbamoyl-D-amino acid amidohydrolase
Overview
N-Acylamino acid racemase (NAAAR) and N-carbamoyl-D-amino-acid amidohydrolase (D-NCAase) are important biocatalysts for producing enantiopure alpha-amino acids. NAAAR forms an octameric assembly and displays induced fit movements upon substrate binding, while D-NCAase is a tetramer that does not change conformation in the presence of a ligand. To investigate the effects of introducing potentially stabilizing S-S bridges in these different multimeric enzymes, cysteine residues predicted to form inter or intra-subunit disulfide bonds were introduced by site-directed mutagenesis. Inter-subunit S-S bonds were formed in two NAAAR variants (A68C-D72C and P60C-Y100C) and two d-NCAase variants (A302C and P295C-F304C). Intra-subunit S-S bonds were formed in two additional NAAAR variants (E149C-A182C and V265C). Crystal structures of NAAARs variants show limited deviations from the wild-type overall tertiary structure. An apo A68C-D72C subunit differs from the wild-type enzyme, in which it has an ordered lid loop, resembling ligand-bound NAAAR. The structures of A222C and A302C D-NCAases are nearly identical to the wild-type enzyme. All mutants with inter-subunit bridges had increases in thermostability. Compared with the wild-type enzyme, A68C-D72C NAAAR showed similar kcat/Km ratios, whereas mutant D-NCAases demonstrated increased kcat/Km ratios at high temperatures (A302C: 4.2-fold at 65 degrees C). Furthermore, molecular dynamic simulations reveal that A302C substantially sustains the fine-tuned catalytic site as temperature increases, achieving enhanced activity.
About this Structure
2GGL is a Single protein structure of sequence from Agrobacterium tumefaciens. This structure supersedes the now removed PDB entries 2FKU and 2BA4. Full crystallographic information is available from OCA.
Reference
Structure-stability-activity relationship in covalently cross-linked N-carbamoyl D-amino acid amidohydrolase and N-acylamino acid racemase., Chiu WC, You JY, Liu JS, Hsu SK, Hsu WH, Shih CH, Hwang JK, Wang WC, J Mol Biol. 2006 Jun 9;359(3):741-53. Epub 2006 Apr 18. PMID:16650857
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