4PVH

Phl p 4 N158H variant, a glucose dehydrogenase

Summary for 4PVH

• Entry DOI: 10.2210/pdb4pvh/pdb
• Related: 4PVE 4PVJ 4PVK 4PWB 4PWC 4PZF
• Descriptor: Pollen allergen Phl p 4.0202, FLAVIN-ADENINE DINUCLEOTIDE, SODIUM ION, ... (5 entities in total)
• Functional Keywords: flavoprotein, bi-covalent flavinylation, oxidoreductase, allergen, glucose dehydrogenase, low oxygen reactivity, grass pollen
• Biological source: Phleum pratense (timothy)
• Total number of polymer chains: 1
• Total formula weight: 56790.37

Authors

Zafred, D.,Teufelberger, A.,Keller, W.,Macheroux, P. (deposition date: 2014-03-17, release date: 2014-04-02, Last modification date: 2024-10-30)

Primary citation

Zafred, D.,Steiner, B.,Teufelberger, A.R.,Hromic, A.,Karplus, P.A.,Schofield, C.J.,Wallner, S.,Macheroux, P.
Rationally engineered flavin-dependent oxidase reveals steric control of dioxygen reduction.
Febs J., 282:3060-3074, 2015

PubMed Abstract: The ability of flavoenzymes to reduce dioxygen varies greatly, and is controlled by the protein environment, which may cause either a rapid reaction (oxidases) or a sluggish reaction (dehydrogenases). Previously, a 'gatekeeper' amino acid residue was identified that controls the reactivity to dioxygen in proteins from the vanillyl alcohol oxidase superfamily of flavoenzymes. We have identified an alternative gatekeeper residue that similarly controls dioxygen reactivity in the grass pollen allergen Phl p 4, a member of this superfamily that has glucose dehydrogenase activity and the highest redox potential measured in a flavoenzyme. A substitution at the alternative gatekeeper site (I153V) transformed the enzyme into an efficient oxidase by increasing dioxygen reactivity by a factor of 60,000. An inverse exchange (V169I) in the structurally related berberine bridge enzyme (BBE) decreased its dioxygen reactivity by a factor of 500. Structural and biochemical characterization of these and additional variants showed that our model enzymes possess a cavity that binds an anion and resembles the 'oxyanion hole' in the proximity of the flavin ring. We showed also that steric control of access to this site is the most important parameter affecting dioxygen reactivity in BBE-like enzymes. Analysis of flavin-dependent oxidases from other superfamilies revealed similar structural features, suggesting that dioxygen reactivity may be governed by a common mechanistic principle.

PubMed: 25619330
DOI: 10.1111/febs.13212

Experimental method

X-RAY DIFFRACTION (1.4 Å)