Protein/Protein and Quantum Dot/Protein Organization in Sequential Monolayer Materials Studied Using Resonance Energy Transfer

Abstract

Controlled junctions of proteins and nanomaterials offer multiple potential applications in the further construction of nanobiodevices. One of the possible junction types is a set of sequential monolayers of various components deposited on a given substrate. The advantage of such an organization is its high sensitivity, resulting from a huge surface covered by molecules or particles. What is more, the molecules/particles adsorbed on a substrate might be easier to handle than the assay in a cuvette. For further application, there should be crosstalk between monolayers; this is defined by the type of individuals forming a complex system. Here, we are studying, using mainly confocal microscopy and FLIM imaging, crosstalk through resonance energy transfer. The sequential monolayers of fluorescent proteins and CdTe quantum dots were deposited on a convenient substrate, a polyvinylidene difluoride membrane. First, we found that the degree of coverage is lower in the second monolayer. Hence, by manipulating the order of deposition, we obtained a system with a varied yield of resonance energy transfer with a donor excess or an acceptor excess. For a deeper understanding of the energy transfer and its limitations in this system influencing the assay pursuit, we utilized Monte Carlo computation. We found that, indeed, the distance between the monolayers, as well as the degree of coverage, is crucial. With the results of the simulation, we might estimate the relative degree of coverage in our sequential monolayers. We also found that in quantum-dots/protein-composed systems, the yield is stronger than predicted by Monte Carlo simulation. Hence, there should be protein reorientation on the nanoparticle surface, leading to such an effect. Finally, we showed that the yield of resonance energy transfer may be modulated by the external application of poly-L-lysines. These chemicals influenced QD fluorescence but not protein fluorescence and might be used, therefore, as a trigger or a switch in nanobiodevices employing those types of sequential monolayers.

Keywords

• monolayers
• fluorescent proteins
• quantum dots
• FRET
• poly-L-lysines