Synthetic Self-Localizing Ligands That Control the Spatial Location of Proteins in Living Cells

Abstract

Small-molecule ligands that control the spatial location of proteins in living cells would be valuable tools for regulating biological systems. However, the creation of such molecules remains almost unexplored because of the lack of a design methodology. Here we introduce a conceptually new type of synthetic ligands, self-localizing ligands (SLLs), which spontaneously localize to specific subcellular regions in mammalian cells. We show that SLLs bind their target proteins and relocate (tether) them rapidly from the cytoplasm to their targeting sites, thus serving as synthetic protein translocators. SLL-induced protein translocation enables us to manipulate diverse synthetic/endogenous signaling pathways. The method is also applicable to reversible protein translocation and allows control of multiple proteins at different times and locations in the same cell. These results demonstrate the usefulness of SLLs in the spatial (and temporal) control of intracellular protein distribution and biological processes, opening a new direction in the design of small-molecule tools or drugs for cell regulation.

A Rationally Designed Upconversion Nanoprobe for in Vivo Detection of Hydroxyl Radical

“…Considering the nondestructive and visual features, fluorescence probe has been established as one of the most useful tools for realtime monitoring chemical species in live cells, tissues and animals. [18][19][20][21][22] Indeed, several elegant fluorescence probes for ·OH have previously been reported using either smallmolecule or nanomaterial fluorophores. [23][24][25][26][27] Nonetheless, the use of these probes is somewhat limited due to the short excitation wavelengths in the UV-vis range, which suffers from shallow tissue penetration depths.…”

SNAP-Tag-Reactive Lipid Anchors Enable Targeted and Spatiotemporally Controlled Localization of Proteins to Phospholipid Membranes

“…Having this capability is essential for the reconstitution of natural membrane protein function and the design of new artificial systems in selfcontained minimal cells. While there have been elegant demonstrations using spontaneous incorporation of certain integral membrane proteins, 12 and reconstitution of the Sec translocon to create lipid/protein membranes, 39 the use of SNAP-tag reactive lipids could offer a simple method to genetically target a much wider range of proteins to the surrounding lipid bilayer ( Figure 3A). We therefore examined whether DSPE-PEG45-BG lipids could be utilized in con- junction with cell free expression systems to genetically trigger membrane localization of a SNAP-tag fusion protein.…”

Ligand-directed dibromophenyl benzoate chemistry for rapid and selective acylation of intracellular natural proteins

“…10–12 Accordingly, we designed four types of new reagents 1–4 containing a TMP ligand and a synthetic fluorophore (7-diethylaminocoumarin (Dc)), connected with ortho -substituted phenyl ester linkers, such as 3,5-dichloro-, 3-nitro-, 3,5-dibromo- and 2,3,5,6-tetrafluoro-4-hydroxy-benzoic acid (Fig. 1c).…”