1CC4

PHE161 AND ARG166 VARIANTS OF P-HYDROXYBENZOATE HYDROXYLASE. IMPLICATIONS FOR NADPH RECOGNITION AND STRUCTURAL STABILITY.

1CC4 の概要

• エントリーDOI: 10.2210/pdb1cc4/pdb
• 分子名称: PROTEIN (P-HYDROXYBENZOATE HYDROXYLASE), FLAVIN-ADENINE DINUCLEOTIDE, P-HYDROXYBENZOIC ACID, ... (4 entities in total)
• 機能のキーワード: oxidoreductase, hydroxybenzoate
• 由来する生物種: Pseudomonas fluorescens
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 45212.08

構造登録者

Eppink, M.H.M.,Bunthof, C.,Schreuder, H.A.,Van Berkel, W.J.H. (登録日: 1999-03-04, 公開日: 1999-03-12, 最終更新日: 2023-08-09)

主引用文献

Eppink, M.H.,Bunthol, C.,Schreuder, H.A.,van Berkel, W.J.
Phe161 and Arg166 variants of p-hydroxybenzoate hydroxylase. Implications for NADPH recognition and structural stability.
Febs Lett., 443:251-255, 1999

PubMed Abstract: Phe161 and Arg166 of p-hydroxybenzoate hydroxylase from Pseudomonas fluorescens belong to a newly discovered sequence motif in flavoprotein hydroxylases with a putative dual function in FAD and NADPH binding [1]. To study their role in more detail, Phe161 and Arg166 were selectively changed by site-directed mutagenesis. F161A and F161G are catalytically competent enzymes having a rather poor affinity for NADPH. The catalytic properties of R166K are similar to those of the native enzyme. R166S and R166E show impaired NADPH binding and R166E has lost the ability to bind FAD. The crystal structure of substrate complexed F161A at 2.2 A is indistinguishable from the native enzyme, except for small changes at the site of mutation. The crystal structure of substrate complexed R166S at 2.0 A revealed that Arg166 is important for providing an intimate contact between the FAD binding domain and a long excursion of the substrate binding domain. It is proposed that this interaction is essential for structural stability and for the recognition of the pyrophosphate moiety of NADPH.

PubMed: 10025942
DOI: 10.1016/S0014-5793(98)01726-8

実験手法

X-RAY DIFFRACTION (2 Å)