Sökning: (WFRF:(Holmér Ingvar)) lar1:(ltu)
> (2000-2004) >
Structural basis fo...
Structural basis for substrate binding and regioselective oxidation of monosaccharides at C3 by pyranose 2-oxidase
-
Kujawa, Magdalena (författare)
-
Ebner, Heidemarie (författare)
-
Leitner, Christian (författare)
-
visa fler...
-
- Hallberg, B. Martin (författare)
- Karolinska Institutet
-
Prongjit, Methinee (författare)
-
Sucharitakul, Jeerus (författare)
-
Ludwig, Roland (författare)
-
Rudsander, Ulla (författare)
-
Peterbauer, Clemens (författare)
-
Chaiyen, Pimchai (författare)
-
Haltrich, Dietmar (författare)
-
- Divne, Christina (författare)
- KTH,Glykovetenskap
-
visa färre...
-
(creator_code:org_t)
- 2006
- 2006
- Engelska.
-
Ingår i: Journal of Biological Chemistry. - 0021-9258 .- 1083-351X. ; 281:46, s. 35104-35115
- Relaterad länk:
-
https://urn.kb.se/re...
-
visa fler...
-
https://doi.org/10.1...
-
http://kipublication...
-
visa färre...
Abstract
Ämnesord
Stäng
- Pyranose2-oxidase(P2Ox) participates in fungal lignin degradation by producing the H2O2 needed for lignin-degrading peroxidases. The enzyme oxidizes cellulose- and hemicellulose-derived aldopyranoses at C2 preferentially, but also on C3, to the corresponding ketoaldoses. To investigate the structural determinants of catalysis, covalent flavinylation, substrate binding, and regios-electivity, wild-type and mutant P2Ox enzymes were produced and characterized biochemically and structurally. Removal of the histidyl-FAD linkage resulted in a catalytically competent enzyme containing tightly, but noncovalently bound FAD. This mutant (H167A) is characterized by a 5-fold lower k(cat), and a 35-mV lower redox potential, although no significant structural changes were seen in its crystal structure. In previous structures of P2Ox, the substrate loop (residues 452-457) covering the active site has been either disordered or in a conformation incompatible with carbohydrate binding. We present here the crystal structure of H167A in complex with a slow substrate, 2-fluoro-2-deoxy-D-glucose. Based on the details of 2-fluoro-2-deoxy-D-glucose binding in position for oxidation at C3, we also outline a probable binding mode for D-glucose positioned for regioselective oxidation at C2. The tentative determinant for discriminating between the two binding modes is the position of the O6 hydroxyl group, which in the C2-oxidation mode can make favorable interactions with Asp(452) in the substrate loop and, possibly, a nearby arginine residue (Arg(472)). We also substantiate our hypothesis with steady-state kinetics data for the alanine replacements of Asp(452) and Arg(472) as well as the double alanine 452/472 mutant.
Nyckelord
- electron-transfer flavoprotein
- p-cresol methylhydroxylase
- fungus trametes-multicolor
- crystal-structure
- phanerochaete-chrysosporium
- cellobiose dehydrogenase
- alcohol oxidase
- covalent flavinylation
- catalytic mechanism
- lignin degradation
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
Hitta via bibliotek
Till lärosätets databas
- Av författaren/redakt...
-
Kujawa, Magdalen ...
-
Ebner, Heidemari ...
-
Leitner, Christi ...
-
Hallberg, B. Mar ...
-
Prongjit, Methin ...
-
Sucharitakul, Je ...
-
visa fler...
-
Ludwig, Roland
-
Rudsander, Ulla
-
Peterbauer, Clem ...
-
Chaiyen, Pimchai
-
Haltrich, Dietma ...
-
Divne, Christina
-
visa färre...
- Artiklar i publikationen
-
Journal of Biolo ...
- Av lärosätet
-
Kungliga Tekniska Högskolan
-
Karolinska Institutet