AI Insight
Researchers developed BOLT (Biotin-Operated Ligand Trapping), a new chemical biology tool that allows scientists to dynamically control where proteins are located inside living cells. The technique uses a HaloTag attached to the protein of interest, streptavidin as a molecular trap, and specially designed fluorescent molecules that can both visualize the protein and either trap it in specific locations or release it on demand using biotin. The team demonstrated BOLT's versatility by controlling secretory cargo release, directing organelle movement along microtubules, relocating nuclear proteins, and tracking both recycled and newly made enzymes moving between the endoplasmic reticulum and Golgi apparatus.
Why it matters
This tool enables researchers to precisely manipulate and track protein locations in living cells with reversible control, which is valuable for understanding how protein location affects function, studying cellular trafficking mechanisms, and potentially developing new therapeutic approaches that depend on controlling where proteins operate within cells.
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⚠️ Preprint – Noch nicht peer-reviewed
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Controlling protein localization through chemical biology tools is essential for mechanistic interrogations, functional perturbations, and therapeutic development. While existing tools cover a wide range of applications, no single approach simultaneously enables inducible trapping, controlled release, and visualization of a protein of interest (POI). To address this gap, we developed Biotin-Operated Ligand Trapping (BOLT), which combines a HaloTag-fused POI with a streptavidin trap. Desthiobiotin-conjugated Janelia Fluor HaloTag ligands enable POI visualization, trapping, and biotin-operated release. We demonstrate BOLT’s capacity in synchronized secretory cargo release, microtubule-directed organelle movement and hitchhiking, and ectopically localizing nuclear transcription factor. Importantly, pulse-chase BOLT further reveals bidirectional endoplasmic reticulum-Golgi trafficking of an endogenous Golgi enzyme and distinguishes recycled from newly synthesized enzyme populations using distinct HTLs. Altogether, BOLT provides an all-in-one platform to interrogate how subcellular localization regulates protein function across HaloTag-compatible systems.