4DUA

cytochrome P450 BM3h-9D7 MRI sensor, no ligand

4DUA の概要

• エントリーDOI: 10.2210/pdb4dua/pdb
• 関連するPDBエントリー: 4DTW 4DTY 4DTZ 4DU2 4DUB 4DUC 4DUD 4DUE 4DUF
• 分子名称: cytochrome P450 BM3 variant 9D7, PROTOPORPHYRIN IX CONTAINING FE, TRIETHYLENE GLYCOL, ... (4 entities in total)
• 機能のキーワード: cytochrome p450, mri contrast sensor, directed evolution, oxidoreductase
• 由来する生物種: Bacillus megaterium
• 細胞内の位置: Cytoplasm : P14779
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 109081.64

構造登録者

Brustad, E.M.,Lelyveld, V.S.,Snow, C.D.,Crook, N.,Martinez, F.M.,Scholl, T.J.,Jasanoff, A.,Arnold, F.H. (登録日: 2012-02-21, 公開日: 2012-06-13, 最終更新日: 2023-09-13)

主引用文献

Brustad, E.M.,Lelyveld, V.S.,Snow, C.D.,Crook, N.,Jung, S.T.,Martinez, F.M.,Scholl, T.J.,Jasanoff, A.,Arnold, F.H.
Structure-guided directed evolution of highly selective p450-based magnetic resonance imaging sensors for dopamine and serotonin.
J.Mol.Biol., 422:245-262, 2012

PubMed Abstract: New tools that allow dynamic visualization of molecular neural events are important for studying the basis of brain activity and disease. Sensors that permit ligand-sensitive magnetic resonance imaging (MRI) are useful reagents due to the noninvasive nature and good temporal and spatial resolution of MR methods. Paramagnetic metalloproteins can be effective MRI sensors due to the selectivity imparted by the protein active site and the ability to tune protein properties using techniques such as directed evolution. Here, we show that structure-guided directed evolution of the active site of the cytochrome P450-BM3 heme domain produces highly selective MRI probes with submicromolar affinities for small molecules. We report a new, high-affinity dopamine sensor as well as the first MRI reporter for serotonin, with which we demonstrate quantification of neurotransmitter release in vitro. We also present a detailed structural analysis of evolved cytochrome P450-BM3 heme domain lineages to systematically dissect the molecular basis of neurotransmitter binding affinity, selectivity, and enhanced MRI contrast activity in these engineered proteins.

PubMed: 22659321
DOI: 10.1016/j.jmb.2012.05.029

実験手法

X-RAY DIFFRACTION (2 Å)