Kinesin-1 activity recorded in living cells with a precipitating dye.
- Resource Type
- Academic Journal
- Authors
- Angerani S; School of Chemistry and Biochemistry, NCCR Chemical Biology, Faculty of Science, University of Geneva, Geneva, Switzerland.; Lindberg E; School of Chemistry and Biochemistry, NCCR Chemical Biology, Faculty of Science, University of Geneva, Geneva, Switzerland.; Klena N; Department of Cell Biology, Faculty of Science, University of Geneva, Geneva, Switzerland.; Bleck CKE; Electron Microscopy Core Facility, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, MD, USA.; Aumeier C; School of Chemistry and Biochemistry, NCCR Chemical Biology, Faculty of Science, University of Geneva, Geneva, Switzerland. charlotte.aumeier@unige.ch.; Winssinger N; School of Chemistry and Biochemistry, NCCR Chemical Biology, Faculty of Science, University of Geneva, Geneva, Switzerland. nicolas.winssinger@unige.ch.
- Source
- Publisher: Nature Pub. Group Country of Publication: England NLM ID: 101528555 Publication Model: Electronic Cited Medium: Internet ISSN: 2041-1723 (Electronic) Linking ISSN: 20411723 NLM ISO Abbreviation: Nat Commun Subsets: MEDLINE
- Subject
- Language
- English
Kinesin-1 is a processive motor protein that uses ATP-derived energy to transport a variety of intracellular cargoes toward the cell periphery. The ability to visualize and monitor kinesin transport in live cells is critical to study the myriad of functions associated with cargo trafficking. Herein we report the discovery of a fluorogenic small molecule substrate (QPD-OTf) for kinesin-1 that yields a precipitating dye along its walking path on microtubules (MTs). QPD-OTf enables to monitor native kinesin-1 transport activity in cellulo without external modifications. In vitro assays show that kinesin-1 and MTs are sufficient to yield fluorescent crystals; in cells, kinesin-1 specific transport of cargo from the Golgi appears as trails of fluorescence over time. These findings are further supported by docking studies, which suggest the binding of the activity-based substrate in the nucleotide binding site of kinesin-1.