Alexander N. Semyonov

Single-molecule motions of oligoarginine transporter conjugates on the plasma membrane of Chinese hamster ovary cells

To explore the real-time dynamic behavior of molecular transporters of the cell-penetrating-peptide (CPP) type on a biological membrane, single fluorescently labeled oligoarginine conjugates were imaged interacting with the plasma membrane of Chinese hamster ovary (CHO) cells. The diffusional motion on the membrane, characterized by single-molecule diffusion coefficient and residence time (tau R), defined as the time from the initial appearance of a single-molecule spot on the membrane (from the solution) to the time the single molecule disappears from the imaging focal plane, was observed for a fluorophore-labeled octaarginine (a model guanidinium-rich CPP) and compared with the corresponding values observed for a tetraarginine conjugate (negative control), a lipid analogue, and a fluorescently labeled protein conjugate (transferrin-Alexa594) known to enter the cell through endocytosis. Imaging of the oligoarginine conjugates was enabled by the use of a new high-contrast fluorophore in the dicyanomethylenedihydrofuran family, which brightens upon interaction with the membrane at normal oxygen concentrations. Taken as a whole, the motions of the octaarginine conjugate single molecules are highly heterogeneous and cannot be described as Brownian motion with a single diffusion coefficient. The observed behavior is also different from that of lipids, known to penetrate cellular membranes through passive diffusion, conventionally involving lateral diffusion followed by membrane bilayer flip-flop. Furthermore, while the octaarginine conjugate behavior shares some common features with transferrin uptake (endocytotic) processes, the two systems also exhibit dissimilar traits when diffusional motions and residence times of single constructs are compared. Additionally, pretreatment of cells with cytochalasin D, a known actin filament disruptor, produces no significant effect, which further rules out unimodal endocytosis as the mechanism of uptake. Also, the involvement of membrane potential in octaarginine-membrane interaction is supported by significant changes in the motion with high [K(+)] treatment. In sum, this first study of single transporter motion on the membrane of a living cell indicates that the mode by which the octaarginine transporter penetrates the cell membrane appears to either be a multimechanism uptake process or a mechanism different from unimodal passive diffusion or endocytosis.

Effect of mobile ions on charge mobility measurements

We present hole time-of-flight (TOF) data on samples of the discotic liquid crystal hexapentyloxytriphenylene containing ionic impurities. By photogenerating holes at various times after reversing the potential across the cell, we have studied the effect of the ions on the measured TOF for various ionic spatial configurations. Comparing the results with simulations of coupled ionic/hole transport, we address the question of whether reliable charge transport data can be extracted for impure samples. Alternatively, we show how TOF experiments provide a useful probe of the spatiotemporal evolution of ionic charge densities under an applied potential.

Interfacial trapping in an aged discotic liquid crystal semiconductor

This study reports on time-of-flight (TOF) hole mobility measurements in aged 2,3,6,7,10,11-Hexakis(pentyloxy)triphenylene columnar liquid crystals. In contrast to the original samples reported in 2006, homeotropically aligned samples yielded TOF transients with an extended non-exponential rise. The experimental data were fit to a simple model that accurately reproduces the TOF transients assuming delayed charge release from traps near the optically excited electrode. While interfacial trapping appears only in the aged materials, the bulk mobility is similar to the pristine material. The model addresses dispersive transport in quasi-one-dimensional materials, determines the charge carrier mobility in systems with interfacial traps, and provides a method for characterizing the traps.

Single crystal, liquid crystal, and hybrid organic semiconductors

The synthesis and characterization of organic semiconductors is being pursued in three primary structure formats: single crystal, liquid crystal and organic-inorganic hybrid. The strategy here is to share common structures, synthesis methods and fabrication techniques across these formats and to utilize common characterization tools such as the time of flight technique. The single crystal efforts concentrate on aromatic and heteroaromatic compounds including simple benzene derivatives and derivatives of the acenes. The structure-property relationships due to incorporation of small substituents and heteroatoms are being examined. Crystals are grown by solution, melt or vapor transport techniques. The liquid crystal studies exploit their self-organizing properties and relative ease of sample preparation. Though calamitic systems tha deliver the largest mobilities are higher order smectics, even some unusual twist grain boundary phases are being studied. We are attempting to synthesize discotic acene derivatives with appropriate substitution patterns to render them mesogenic. The last format being examined is the hybrid organic-inorganic class. Here, layered materials of alternating organic and inorganic composition are designed and synthesized. Typical materials are conjugated aromatic compounds, usually functinalized with an amine or a pyridine and reacted with appropriate reactive metal derivatives to incorporate them into metal oxide or sulfide layers.