Confirmed via MRI contrast imaging Testing of Variants In Vitro In Vivo PC12 cells- derived from adrenal gland of rats-in F12-K
Dopamine release only in presence of extracellular K + Brain of anesthetized rats Conclusions Created MRI contrast agents sensitive to neurotransmitter dopamine heme domain of flavocytochrome BM3
Developed novel screening methodology - ligand induced decrease in relaxivity under MRI -and optical characterization
Evolved specificity of BM3h-based sensors away from natural ligand and toward dopamine via error-prone PCR.
Introduced mutation I366V by site-directed mutagenesis to enhance thermostability and tolerance to further mutation
Performed In Vitro (PC12) and In Vivo (live animal) testing of efficacy of developed sensors
General paradigm for development of molecular probes for MRI with tunable ligand-binding and catalytic properties