4DUC

cytochrome P450 BM3h-2G9 MRI sensor, no ligand

Summary for 4DUC

• Entry DOI: 10.2210/pdb4duc/pdb
• Related: 4DTW 4DTY 4DTZ 4DU2 4DUA 4DUB 4DUD 4DUE 4DUF
• Descriptor: cytochrome P450 BM3 variant 2G9, PROTOPORPHYRIN IX CONTAINING FE (3 entities in total)
• Functional Keywords: cytochrome p450, mri contrast sensor, directed evolution, oxidoreductase
• Biological source: Bacillus megaterium
• Cellular location: Cytoplasm (By similarity): P14779
• Total number of polymer chains: 2
• Total formula weight: 109189.98

Authors

Brustad, E.M.,Lelyveld, V.S.,Snow, C.D.,Crook, N.,Martinez, F.M.,Scholl, T.J.,Jasanoff, A.,Arnold, F.H. (deposition date: 2012-02-21, release date: 2012-06-13, Last modification date: 2023-09-13)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (1.92 Å)