2VVM

The structure of MAO-N-D5, a variant of monoamine oxidase from Aspergillus niger.

2VVM の概要

• エントリーDOI: 10.2210/pdb2vvm/pdb
• 関連するPDBエントリー: 2VVL
• 分子名称: MONOAMINE OXIDASE N, FLAVIN-ADENINE DINUCLEOTIDE, PROLINE, ... (5 entities in total)
• 機能のキーワード: monoamine oxidase, aspergillus niger, fad, peroxisome, flavoprotein, oxidoreductase, enantioselectivity, directed evolution variant
• 由来する生物種: ASPERGILLUS NIGER
• 細胞内の位置: Peroxisome (Probable): P46882
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 113324.67

構造登録者

Atkin, K.E.,Hart, S.,Turkenburg, J.P.,Brzozowski, A.M.,Grogan, G.J. (登録日: 2008-06-10, 公開日: 2008-11-04, 最終更新日: 2024-05-01)

主引用文献

Atkin, K.E.,Reiss, R.,Koehler, V.,Bailey, K.R.,Hart, S.,Turkenburg, J.P.,Turner, N.J.,Brzozowski, A.M.,Grogan, G.J.
The Structure of Monoamine Oxidase from Aspergillus Niger Provides a Molecular Context for Improvements in Activity Obtained by Directed Evolution.
J.Mol.Biol., 384:1218-, 2008

PubMed Abstract: Monoamine oxidase from Aspergillus niger (MAO-N) is a flavoenzyme that catalyses the oxidative deamination of primary amines. MAO-N has been used as the starting model for a series of directed evolution experiments, resulting in mutants of improved activity and broader substrate specificity, suitable for application in the preparative deracemisation of primary, secondary and tertiary amines when used as part of a chemoenzymatic oxidation-reduction cycle. The structures of a three-point mutant (Asn336Ser/Met348Lys/Ile246Met or MAO-N-D3) and a five-point mutant (Asn336Ser/Met348Lys/Ile246Met/Thr384Asn/Asp385Ser or MAO-N-D5) have been obtained using a multiple-wavelength anomalous diffraction experiment on a selenomethionine derivative of the truncated MAO-N-D5 enzyme. MAO-N exists as a homotetramer with a large channel at its centre and shares some structural features with human MAO B (MAO-B). A hydrophobic cavity extends from the protein surface to the active site, where a non-covalently bound flavin adenine dinucleotide (FAD) sits at the base of an 'aromatic cage,' the sides of which are formed by Trp430 and Phe466. A molecule of l-proline was observed near the FAD, and this ligand superimposed well with isatin, a reversible inhibitor of MAO-B, when the structures of MAO-N proline and MAO-B-isatin were overlaid. Of the mutations that confer the ability to catalyse the oxidation of secondary amines in MAO-N-D3, Asn336Ser reduces steric bulk behind Trp430 of the aromatic cage and Ile246Met confers greater flexibility within the substrate binding site. The two additional mutations, Thr384Asn and Asp385Ser, that occur in the MAO-N-D5 variant, which is able to oxidise tertiary amines, appear to influence the active-site environment remotely through changes in tertiary structure that perturb the side chain of Phe382, again altering the steric and electronic character of the active site near FAD. The possible implications of the change in steric and electronic environment caused by relevant mutations are discussed with respect to the improved catalytic efficiency of the MAO-N variants described in the literature.

PubMed: 18951902
DOI: 10.1016/J.JMB.2008.09.090

実験手法

X-RAY DIFFRACTION (1.85 Å)