- Metal trafficking in neurons: protein interaction dynamics
Metal trafficking in neurons is mediated by protein-protein (P-P) interactions between metalloproteins. Choosing Atox1/ATP7A, together with CCS/SOD1, as model Cu chaperone-transporter systems, we study P-P interaction dynamics at single-molecule level. We quantitatively determine cellular protein concentration, spatial distribution, diffusive behavior, interaction dynamics and thermodynamics simultaneously under different Cu stressed conditions in both healthy and diseased neurons.
- Metal trafficking at synapses: effect of synaptic Cu
Choosing Atox1/ATP7A as the synaptic Cu release system; we examine Cu release mechanisms into the synaptic cleft. With genetically introduce a SNAP tag to a synaptic protein (e.g., N-methyl-D-aspartate receptor, NMDAR), followed by covalently attaching a Cu sensor. We quantitatively determine metal release time in single synapses by analyzing fluorescence images of tagged fluorescent protein and sensor.