Structure-guided directed evolution of highly selective P450-based magnetic resonance imaging sensors for dopamine and serotonin

Eric M. Brustad, Victor S. Lelyveld, Christopher D. Snow, Nathan Crook, Sang Taek Jung, Francisco M. Martinez, Timothy J. Scholl, Alan Jasanoff, Frances H. Arnold

Research output: Contribution to journalArticle

25 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)245-262
Number of pages18
JournalJournal of Molecular Biology
Volume422
Issue number2
DOIs
Publication statusPublished - 2012 Sep 14
Externally publishedYes

Fingerprint

Dopamine
Serotonin
Magnetic Resonance Imaging
Heme
Cytochrome P-450 Enzyme System
Neurotransmitter Agents
Catalytic Domain
Metalloproteins
Proteins
Brain Diseases
Molecular Dynamics Simulation
Ligands

Keywords

  • biosensors
  • cytochrome P450
  • directed evolution
  • MRI contrast agents
  • neurotransmitter

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Structure-guided directed evolution of highly selective P450-based magnetic resonance imaging sensors for dopamine and serotonin. / Brustad, Eric M.; Lelyveld, Victor S.; Snow, Christopher D.; Crook, Nathan; Jung, Sang Taek; Martinez, Francisco M.; Scholl, Timothy J.; Jasanoff, Alan; Arnold, Frances H.

In: Journal of Molecular Biology, Vol. 422, No. 2, 14.09.2012, p. 245-262.

Research output: Contribution to journalArticle

Brustad, EM, Lelyveld, VS, Snow, CD, Crook, N, Jung, ST, Martinez, FM, Scholl, TJ, Jasanoff, A & Arnold, FH 2012, 'Structure-guided directed evolution of highly selective P450-based magnetic resonance imaging sensors for dopamine and serotonin', Journal of Molecular Biology, vol. 422, no. 2, pp. 245-262. https://doi.org/10.1016/j.jmb.2012.05.029
Brustad, Eric M. ; Lelyveld, Victor S. ; Snow, Christopher D. ; Crook, Nathan ; Jung, Sang Taek ; Martinez, Francisco M. ; Scholl, Timothy J. ; Jasanoff, Alan ; Arnold, Frances H. / Structure-guided directed evolution of highly selective P450-based magnetic resonance imaging sensors for dopamine and serotonin. In: Journal of Molecular Biology. 2012 ; Vol. 422, No. 2. pp. 245-262.
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