1e8g: Difference between revisions
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[[Image:1e8g.gif|left|200px]]<br /> | [[Image:1e8g.gif|left|200px]]<br /><applet load="1e8g" size="450" color="white" frame="true" align="right" spinBox="true" | ||
<applet load="1e8g" size="450" color="white" frame="true" align="right" spinBox="true" | |||
caption="1e8g, resolution 2.10Å" /> | caption="1e8g, resolution 2.10Å" /> | ||
'''STRUCTURE OF THE H61T DOUBLE MUTANT OF VANILLYL-ALCOHOL OXIDASE IN COMPLEX WITH FLUORO-CRESOL'''<br /> | '''STRUCTURE OF THE H61T DOUBLE MUTANT OF VANILLYL-ALCOHOL OXIDASE IN COMPLEX WITH FLUORO-CRESOL'''<br /> | ||
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==About this Structure== | ==About this Structure== | ||
1E8G is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Penicillium_simplicissimum Penicillium simplicissimum] with FAD and FCR as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Vanillyl-alcohol_oxidase Vanillyl-alcohol oxidase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.1.3.38 1.1.3.38] | 1E8G is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/Penicillium_simplicissimum Penicillium simplicissimum] with FAD and FCR as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Vanillyl-alcohol_oxidase Vanillyl-alcohol oxidase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.1.3.38 1.1.3.38] Known structural/functional Sites: <scene name='pdbsite=AC1:Fad Binding Site For Chain A'>AC1</scene>, <scene name='pdbsite=AC2:Fcr Binding Site For Chain A'>AC2</scene>, <scene name='pdbsite=AC3:Fad Binding Site For Chain B'>AC3</scene> and <scene name='pdbsite=AC4:Fcr Binding Site For Chain B'>AC4</scene>. Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1E8G OCA]. | ||
==Reference== | ==Reference== | ||
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[[Category: specificity]] | [[Category: specificity]] | ||
''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on | ''Page seeded by [http://ispc.weizmann.ac.il/oca OCA ] on Tue Dec 18 15:03:58 2007'' | ||
Revision as of 15:54, 18 December 2007
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STRUCTURE OF THE H61T DOUBLE MUTANT OF VANILLYL-ALCOHOL OXIDASE IN COMPLEX WITH FLUORO-CRESOL
OverviewOverview
Vanillyl-alcohol oxidase (VAO) is member of a newly recognized, flavoprotein family of structurally related oxidoreductases. The enzyme, contains a covalently linked FAD cofactor. To study the mechanism of, flavinylation we have created a design point mutation (His-61 --> Thr). In, the mutant enzyme the covalent His-C8alpha-flavin linkage is not formed, while the enzyme is still able to bind FAD and perform catalysis. The H61T, mutant displays a similar affinity for FAD and ADP (K(d) = 1.8 and 2.1, microm, respectively) but does not interact with FMN. H61T is about, 10-fold less active with 4-(methoxymethyl)phenol) (k(cat) = 0.24 s(-)(1), K(m) = 40 microm) than the wild-type enzyme. The crystal structures of, both the holo and apo form of H61T are highly similar to the structure of, wild-type VAO, indicating that binding of FAD to the apoprotein does not, require major structural rearrangements. These results show that covalent, flavinylation is an autocatalytical process in which His-61 plays a, crucial role by activating His-422. Furthermore, our studies clearly, demonstrate that in VAO, the FAD binds via a typical lock-and-key approach, to a preorganized binding site.
About this StructureAbout this Structure
1E8G is a Single protein structure of sequence from Penicillium simplicissimum with FAD and FCR as ligands. Active as Vanillyl-alcohol oxidase, with EC number 1.1.3.38 Known structural/functional Sites: , , and . Full crystallographic information is available from OCA.
ReferenceReference
Structural analysis of flavinylation in vanillyl-alcohol oxidase., Fraaije MW, van Den Heuvel RH, van Berkel WJ, Mattevi A, J Biol Chem. 2000 Dec 8;275(49):38654-8. PMID:10984479
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