1e8f: Difference between revisions
No edit summary |
No edit summary |
||
| Line 1: | Line 1: | ||
[[Image:1e8f.gif|left|200px]] | [[Image:1e8f.gif|left|200px]] | ||
<!-- | |||
The line below this paragraph, containing "STRUCTURE_1e8f", creates the "Structure Box" on the page. | |||
You may change the PDB parameter (which sets the PDB file loaded into the applet) | |||
or the SCENE parameter (which sets the initial scene displayed when the page is loaded), | |||
or leave the SCENE parameter empty for the default display. | |||
| | --> | ||
| | {{STRUCTURE_1e8f| PDB=1e8f | SCENE= }} | ||
}} | |||
'''STRUCTURE OF THE H61T MUTANT OF THE FLAVOENZYME VANILLYL-ALCOHOL OXIDASE IN THE APO FORM''' | '''STRUCTURE OF THE H61T MUTANT OF THE FLAVOENZYME VANILLYL-ALCOHOL OXIDASE IN THE APO FORM''' | ||
| Line 28: | Line 25: | ||
[[Category: Fraaije, M.]] | [[Category: Fraaije, M.]] | ||
[[Category: Mattevi, A.]] | [[Category: Mattevi, A.]] | ||
[[Category: | [[Category: Catalysis]] | ||
[[Category: | [[Category: Flavoenzyme]] | ||
[[Category: | [[Category: Flavoprotein]] | ||
[[Category: | [[Category: Methanol utilization]] | ||
[[Category: | [[Category: Oxidase]] | ||
[[Category: | [[Category: Oxidoreductase]] | ||
[[Category: | [[Category: Peroxisome]] | ||
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Fri May 2 14:47:37 2008'' | |||
''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on | |||
Revision as of 14:47, 2 May 2008
STRUCTURE OF THE H61T MUTANT OF THE FLAVOENZYME VANILLYL-ALCOHOL OXIDASE IN THE APO FORM
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
1E8F is a Single protein structure of sequence from Penicillium simplicissimum. 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 Page seeded by OCA on Fri May 2 14:47:37 2008