Identification and role of ionizing functional groups at the active center of Rhodotorula gracilis D-amino acid oxidase

Pollegioni, Loredano; Harris, Christopher M.; Molla, Gianluca; Pilone, Mirella S.; Ghisla, Sandro

Identification and role of ionizing functional groups at the active center of Rhodotorula gracilis D-amino acid oxidase

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Identification_and_role_of_ionizing_functional_groups_at_the_active_center_of_Rhodotorula_gracilis_D_amino_acid_oxidase.pdf(172.98 KB)

Date

2001

Authors

Pollegioni, Loredano

Harris, Christopher M.

Molla, Gianluca

Pilone, Mirella S.

Ghisla, Sandro

Publication type

Journal article

Published in

FEBS letters ; 507 (2001), 3. - pp. 323-326. - ISSN 0014-5793

Abstract

D-Amino acid oxidase (DAAO) is a flavoprotein oxidase that catalyzes the oxidation of amino acids and produces ketoacids and H2O2. The rate of product release from reduced DAAO from Rhodotorula gracilis is pH dependent and reflects a pKa of ~9.3. Binding of benzoate and 3,3,3-trifluoro-Image-alanine to wild-type and Y238F DAAO is also pH dependent (pKa=9.8±0.1 and 9.05±0.1, respectively for benzoate binding). However, binding of benzoate to Y223F DAAO is pH independent, indicating the pKa is due to Y223 OH. This latter residue is thus involved in substrate binding, and probably is the group that governs product release. In contrast to this, the second active site tyrosine, Y238, has little influence on ligand binding.

Subject (DDC)

570 Biosciences, Biology

Keywords

D-Amino acid oxidase,Flavoprotein,pH effect,Ligand binding,Ionization,Catalytic mechanism

Cite This

ISO 690

POLLEGIONI, Loredano, Christopher M. HARRIS, Gianluca MOLLA, Mirella S. PILONE, Sandro GHISLA, 2001. Identification and role of ionizing functional groups at the active center of Rhodotorula gracilis D-amino acid oxidase. In: FEBS letters. 507(3), pp. 323-326. ISSN 0014-5793. Available under: doi: 10.1016/S0014-5793(01)02983-0

BibTex

@article{Pollegioni2001Ident-7065,
  year={2001},
  doi={10.1016/S0014-5793(01)02983-0},
  title={Identification and role of ionizing functional groups at the active center of Rhodotorula gracilis D-amino acid oxidase},
  number={3},
  volume={507},
  issn={0014-5793},
  journal={FEBS letters},
  pages={323--326},
  author={Pollegioni, Loredano and Harris, Christopher M. and Molla, Gianluca and Pilone, Mirella S. and Ghisla, Sandro}
}

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