Single particle tracking (SPT) provides information about the microscopic motions of individual particles in live cells.
We applied SPT to study the diffusion of membrane transport proteins in cell plasma membranes in which individual
proteins are labeled with quantum dots at engineered extracellular epitopes. Software was created to deduce particle
diffusive modes from quantum dot trajectories. SPT of aquaporin (AQP) water channels and cystic fibrosis
transmembrane conductance regulator (CFTR) chloride channels revealed several types of diffusion. AQP1 was freely
mobile in cell membranes, showing rapid, Brownian-type diffusion. The full-length (M1) isoform of AQP4 also diffused
rapidly, though the diffusion of a shorter (M23) isoform of AQP4 was highly restricted due to its supermolecular
assembly in raft-like orthogonal arrays. CFTR mobility was also highly restricted, in a spring-like potential, due to its
tethering to the actin cytoskeleton through PDZ-domain C-terminus interactions. The biological significance of
regulated diffusion of membrane transport proteins is a subject of active investigation.
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