2vka
From Proteopedia
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| - | [[Image:2vka.jpg|left|200px]] | + | [[Image:2vka.jpg|left|200px]] |
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| - | '''SITE-DIRECTED MUTAGENESIS OF THE CATALYTIC TRYPTOPHAN ENVIRONMENT IN PLEUROTUS ERYNGII VERSATILE PEROXIDASE''' | + | {{Structure |
| + | |PDB= 2vka |SIZE=350|CAPTION= <scene name='initialview01'>2vka</scene>, resolution 2.000Å | ||
| + | |SITE= <scene name='pdbsite=AC1:Hem+Binding+Site+For+Chain+A'>AC1</scene>, <scene name='pdbsite=AC2:So4+Binding+Site+For+Chain+A'>AC2</scene>, <scene name='pdbsite=AC3:Ca+Binding+Site+For+Chain+A'>AC3</scene>, <scene name='pdbsite=AC4:Ca+Binding+Site+For+Chain+A'>AC4</scene> and <scene name='pdbsite=AC5:Gol+Binding+Site+For+Chain+A'>AC5</scene> | ||
| + | |LIGAND= <scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene>, <scene name='pdbligand=CA:CALCIUM+ION'>CA</scene>, <scene name='pdbligand=HEM:PROTOPORPHYRIN+IX+CONTAINING+FE'>HEM</scene> and <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene> | ||
| + | |ACTIVITY= | ||
| + | |GENE= | ||
| + | }} | ||
| + | |||
| + | '''SITE-DIRECTED MUTAGENESIS OF THE CATALYTIC TRYPTOPHAN ENVIRONMENT IN PLEUROTUS ERYNGII VERSATILE PEROXIDASE''' | ||
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==Overview== | ==Overview== | ||
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==About this Structure== | ==About this Structure== | ||
| - | 2VKA is a [ | + | 2VKA is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Pleurotus_eryngii Pleurotus eryngii]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=2VKA OCA]. |
==Reference== | ==Reference== | ||
| - | Site-Directed Mutagenesis of the Catalytic Tryptophan Environment in Pleurotus eryngii Versatile Peroxidase(,)., Ruiz-Duenas FJ, Morales M, Mate MJ, Romero A, Martinez MJ, Smith AT, Martinez AT, Biochemistry. 2008 Feb 12;47(6):1685-95. Epub 2008 Jan 18. PMID:[http:// | + | Site-Directed Mutagenesis of the Catalytic Tryptophan Environment in Pleurotus eryngii Versatile Peroxidase(,)., Ruiz-Duenas FJ, Morales M, Mate MJ, Romero A, Martinez MJ, Smith AT, Martinez AT, Biochemistry. 2008 Feb 12;47(6):1685-95. Epub 2008 Jan 18. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/18201105 18201105] |
[[Category: Pleurotus eryngii]] | [[Category: Pleurotus eryngii]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
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[[Category: allelic variant]] | [[Category: allelic variant]] | ||
[[Category: aromatic-substrate binding]] | [[Category: aromatic-substrate binding]] | ||
| - | [[Category: class ii ( fungal) | + | [[Category: class ii ( fungal) peroxidase]] |
[[Category: electron transfer]] | [[Category: electron transfer]] | ||
[[Category: homology modeling]] | [[Category: homology modeling]] | ||
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[[Category: lignin peroxidase]] | [[Category: lignin peroxidase]] | ||
[[Category: manganese peroxidase]] | [[Category: manganese peroxidase]] | ||
| - | [[Category: mn-independent oxidation phenolic non-phenolic | + | [[Category: mn-independent oxidation phenolic non-phenolic aromatic]] |
[[Category: mnii oxidation]] | [[Category: mnii oxidation]] | ||
[[Category: oxidoreductase]] | [[Category: oxidoreductase]] | ||
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[[Category: polyvalent peroxidase]] | [[Category: polyvalent peroxidase]] | ||
| - | ''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 18:47:23 2008'' |
Revision as of 16:47, 20 March 2008
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| , resolution 2.000Å | |||||||
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| Sites: | , , , and | ||||||
| Ligands: | , , and | ||||||
| Coordinates: | save as pdb, mmCIF, xml | ||||||
SITE-DIRECTED MUTAGENESIS OF THE CATALYTIC TRYPTOPHAN ENVIRONMENT IN PLEUROTUS ERYNGII VERSATILE PEROXIDASE
Overview
Lignin degradation by fungal peroxidases is initiated by one-electron transfer to an exposed tryptophan radical, a reaction mediated by veratryl alcohol (VA) in lignin peroxidase (LiP). Versatile peroxidase (VP) differs not only in its oxidation of Mn2+ at a second catalytic site but also in its ability to directly oxidize different aromatic compounds. The catalytic tryptophan environment was compared in LiP and VP crystal structures, and six residues near VP Trp164 were modified by site-directed mutagenesis. Oxidation of Mn2+ was practically unaffected. However, several mutations modified the oxidation kinetics of the high-redox-potential substrates VA and Reactive Black 5 (RB5), demonstrating that other residues contribute to substrate oxidation by the Trp164 radical. Introducing acidic residues at the tryptophan environment did not increase the efficiency of VP oxidizing VA. On the contrary, all variants harboring the R257D mutation lost their activity on RB5. Interestingly, this activity was restored when VA was added as a mediator, revealing the LiP-type behavior of this variant. Moreover, combination of the A260F and R257A mutations strongly increased (20-50-fold) the apparent second-order rate constants for reduction of VP compounds I and II by VA to values similar to those found in LiP. Dissociation of the enzyme-product complex seemed to be the limiting step in the turnover of this improved variant. Nonexposed residues in the vicinity of Trp164 can also affect VP activity, as found with the M247F mutation. This was a direct effect since no modification of the surrounding residues was found in the crystal structure of this variant.
About this Structure
2VKA is a Single protein structure of sequence from Pleurotus eryngii. Full crystallographic information is available from OCA.
Reference
Site-Directed Mutagenesis of the Catalytic Tryptophan Environment in Pleurotus eryngii Versatile Peroxidase(,)., Ruiz-Duenas FJ, Morales M, Mate MJ, Romero A, Martinez MJ, Smith AT, Martinez AT, Biochemistry. 2008 Feb 12;47(6):1685-95. Epub 2008 Jan 18. PMID:18201105
Page seeded by OCA on Thu Mar 20 18:47:23 2008
Categories: Pleurotus eryngii | Single protein | Martinez, A T. | Martinez, M J. | Mate, M J. | Morales, M. | Romero, A. | Ruiz-Duenas, F J. | Smith, A. | CA | GOL | HEM | SO4 | Allelic variant | Aromatic-substrate binding | Class ii ( fungal) peroxidase | Electron transfer | Homology modeling | Lignin degradation | Lignin peroxidase | Manganese peroxidase | Mn-independent oxidation phenolic non-phenolic aromatic | Mnii oxidation | Oxidoreductase | Peroxidase | Polyvalent peroxidase
