Dr. Glenn Carrington, Biology, Leeds
Kif5a (Kinesin-1) is a microtubule-based motor protein implicated in neurodegeneration. It autoregulates its activity via interaction of the tail with the motor domains. We recently investigated the structure of full-length and mutated autoinhibited KIF5a using a combination of electron-microscopy, cross-linking mass-spectrometry, and molecular modelling to reveal the interactions necessary for autoinhibition. The mutated form has been reported to make kinesin constitutively active, yet we found that auto-inhibition for this mutant was not abolished in vitro. In this project, we will test the alternative possibility that this mutation affects KIF5a’s interaction with binding partners in vivo, which leads to the constitutively active phenotype. To test this, the student will culture and differentiate neuronal cells, and perform pull-down assays to reveal whether the mutant kinesin has altered binding partners compared to wild-type. We will characterise candidate interacting proteins using electron microscopy, mass spectrometry, and mass photometry. In parallel, we will also use in vitro motility assays and live-cell imaging to investigate the motile behaviour of protein mutants. This provides the student with hands-on experience in cell culture and protein work. Students will be expected to present their findings orally at a research day in York in September 2024.