DNA-Programed Plasmon Rulers Decrypt Single-Receptor Dimerization on Cell Membrane

Abstract

Decrypting the dynamics of receptor dimerization on cell membranes bears great importance in identifying the mechanisms regulating diverse cellular activities. In this regard, long-term monitoring of single-molecule behavior during receptor dimerization allows deepening insight into the dimerization process and tracking of the behavior of individual receptors, yet this remains to be realized. Herein, real-time observation of the receptor tyrosine kinases family (RTKs) at single-molecule level based on plasmon rulers was achieved for the first time, which enabled precise regulation and dynamic monitoring of the dimerization process by DNA programming with excellent photostability. Additionally, those nanoprobes demonstrated substantial application in the regulation of RTKs protein dimerization/phosphorylation and activation of downstream signaling pathways.The proposed nanoprobes hold considerable potential for elucidating the molecular mechanisms of single-receptor dimerization as well as the conformational transitions upon dimerization, providing a new paradigm for the precise manipulation and monitoring of specific single-receptor crosslink events in biological systems.

Transformable DNA Nanorobots Reversibly Regulating Cell Membrane Receptors for Modulation of Cellular Migrations

“…Dimerization of the Met receptors on the cell membrane can induce self-phosphorylation and further accelerate cell migration (Figure A) . To verify the feasibility of the designed nR for regulating cell behavior, DU145 cells (a human prostate cancer cell line) with moderate expression of Met were chosen as a model .…”

“…As shown in Figure S6, the cell cytotoxicity experiment revealed tion (Figure 3A). 28 To verify the feasibility of the designed nR for regulating cell behavior, DU145 cells (a human prostate cancer cell line) with moderate expression of Met were chosen as a model. 19 The DNA nanomachines were still working on the surface of the DU145 cells (Figure S7).…”

“…26,27 plasmon rulers that held the potential for elucidating the molecular mechanisms of single-receptor dimerization and conformational transitions upon dimerization. 28 While these ingenious applications of DNA nanotechnology provide infinite possibilities for cellular engineering in biology, 29,30 a strategy that can manipulate cellular function through the simple operation of DNA nanomachines is desirable. However, most of the designed DNA nanodevices can only promote the oligomerization of receptors, not achieve the deoligomerization for reversible regulation.…”

“…Yang et al proposed a bispecific aptamer that worked not only as a blocking assistant to inhibit downstream signaling pathways but also as a cancer biomarker for enhancing cell selectivity . The progression of DNA nanotechnology for controlling cellular behaviors enhances the understanding of the receptor molecule working mechanisms and contributes to cancer therapeutics. , Xu et al reported DNA-programmed plasmon rulers that held the potential for elucidating the molecular mechanisms of single-receptor dimerization and conformational transitions upon dimerization . While these ingenious applications of DNA nanotechnology provide infinite possibilities for cellular engineering in biology, , a strategy that can manipulate cellular function through the simple operation of DNA nanomachines is desirable.…”

Ultrasensitive Multiplex Imaging of Cell Surface Proteins via Core-Shell Surface-Enhanced Raman Scattering Nanoprobes

“…Numerous proteins on the cell membrane surface function as one of the important mediators for the interaction and communication between the cells and their surrounding microenvironment. , Among them, a large proportion of cell surface proteins are specific receptors with highly dynamic expression levels, participating in some important biological processes and events. − Therefore, they are often considered as important biomarkers for disease diagnosis. , For example, human epidermal growth factor receptors (HER2) have been shown to be overexpressed in approximately 20–30% of breast cancer tumors, making them an important biomarker for the diagnosis and treatment of breast cancer. , Rapid monitoring of cell surface protein expression levels as well as their activity status will be of great importance for improving the accuracy and throughput of cell identification and disease diagnosis, which is helpful to obtain rich biological information on how cells respond to external stimuli. , Currently, characterization of cell surface proteins is largely attributed to various cell imaging techniques. − For example, as a well-established and accepted imaging technique, fluorescence analysis has been widely used for the detection of membrane surface proteins. − However, low quantum yields, photobleaching, and interference from cellular autofluorescence in fluorescence imaging are still important obstacles in long-term tracking and high spatial resolution detection . Moreover, owing to the broad bandwidth of the fluorescence spectrum, the overlapping of emission spectra between different fluorescent dyes is unavoidable, which will undoubtedly interfere with multicolor imaging applications for multiplexed detection …”