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TitleStructure of Alcohol Oxidase from Pichia pastoris by Cryo-Electron Microscopy.
Journal, issue, pagesPLoS One, Vol. 11, Issue 7, Page e0159476, Year 2016
Publish dateJul 26, 2016
AuthorsJanet Vonck / David N Parcej / Deryck J Mills /
PubMed AbstractThe first step in methanol metabolism in methylotrophic yeasts, the oxidation of methanol and higher alcohols with molecular oxygen to formaldehyde and hydrogen peroxide, is catalysed by alcohol ...The first step in methanol metabolism in methylotrophic yeasts, the oxidation of methanol and higher alcohols with molecular oxygen to formaldehyde and hydrogen peroxide, is catalysed by alcohol oxidase (AOX), a 600-kDa homo-octamer containing eight FAD cofactors. When these yeasts are grown with methanol as the carbon source, AOX forms large crystalline arrays in peroxisomes. We determined the structure of AOX by cryo-electron microscopy at a resolution of 3.4 Å. All residues of the 662-amino acid polypeptide as well as the FAD are well resolved. AOX shows high structural homology to other members of the GMC family of oxidoreductases, which share a conserved FAD binding domain, but have different substrate specificities. The preference of AOX for small alcohols is explained by the presence of conserved bulky aromatic residues near the active site. Compared to the other GMC enzymes, AOX contains a large number of amino acid inserts, the longest being 75 residues. These segments are found at the periphery of the monomer and make extensive inter-subunit contacts which are responsible for the very stable octamer. A short surface helix forms contacts between two octamers, explaining the tendency of AOX to form crystals in the peroxisomes.
External linksPLoS One / PubMed:27458710 / PubMed Central
MethodsEM (single particle)
Resolution3.37 Å
Structure data

8072

5i68

EMDB-8072, PDB-5i68:
Alcohol oxidase from Pichia pastoris
Method: EM (single particle) / Resolution: 3.37 Å

Chemicals

ChemComp-MG:
Unknown entry

ChemComp-FAD:
FLAVIN-ADENINE DINUCLEOTIDE / FAD*YM

Source
  • komagataella pastoris (fungus)
KeywordsOXIDOREDUCTASE / alcohol oxidase peroxisome

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