A de novo designed coiled coil-based switch regulates the microtubule motor kinesin-1

DOI: 10.1038/s41589-024-01640-2

Authors: Jessica A. Cross (University of Bristol) , William M. Dawson (University of Bristol) , Shivam R. Shukla (University of Bristol) , Johannes F. Weijman (University of Bristol) , Judith Mantell (University of Bristol) , Mark P. Dodding (University of Bristol) , Derek N. Woolfson (University of Bristol)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Nature Chemical Biology , VOL 53

State: Published (Approved)
Published: June 2024
Diamond Proposal Number(s): 37518 , 35672 , 31440

Abstract: Many enzymes are allosterically regulated via conformational change; however, our ability to manipulate these structural changes and control function is limited. Here we install a conformational switch for allosteric activation into the kinesin-1 microtubule motor in vitro and in cells. Kinesin-1 is a heterotetramer that accesses open active and closed autoinhibited states. The equilibrium between these states centers on a flexible elbow within a complex coiled-coil architecture. We target the elbow to engineer a closed state that can be opened with a de novo designed peptide. The alternative states are modeled computationally and confirmed by biophysical measurements and electron microscopy. In cells, peptide-driven activation increases kinesin transport, demonstrating a primary role for conformational switching in regulating motor activity. The designs are enabled by our understanding of ubiquitous coiled-coil structures, opening possibilities for controlling other protein activities.

Diamond Keywords: Enzymes

Subject Areas: Biology and Bio-materials, Chemistry


Instruments: B21-High Throughput SAXS

Added On: 09/06/2024 20:55

Documents:
s41589-024-01640-2.pdf

Discipline Tags:

Biochemistry Chemistry Structural biology Biophysics Life Sciences & Biotech

Technical Tags:

Scattering Small Angle X-ray Scattering (SAXS)