Faculty of Biological Sciences

Molecular Contractility, Molecular Motors and Microscopy Research Group

Research Interests:

We are interested in

Structure and function of the contractile apparatus of muscle; the myofibril,

Structure and functions of the myosin superfamily

Kinesin and dynein.

'Super-resolution imaging'.

We use a wide range of techniques (wet biochemistry (protein isolation, expression and characterisation), biophysical methods (eg analytical ultracentrifugation, atomic force microscopy), light microscopy (confocal, time-lapse fluorescence, deconvolution and super-resolution microscopies) electron microscopy, including time-resolved cryo-EM, molecular genetic tools and cell biology (tissue culture, live cell imaging) to investigate the structure and function of the muscle proteins myosin, titin and actin.

We also investigate cell behaviour, and the dynamic behaviour of molecular motors within cells, using time-lapse phase and fluorescence microscopy and most recently using our new instant structured illumination microscope.  Our research forms part of the major effort in structural biology at Leeds.

picture of lab members

Some of our research group, on a cold frosty January day. (From Left to right: Alistair Curd, Chris Bartlett, Francine Parker, Glenn McConkey, Ruth Hughes, Marcin Wolny, Anna Lopata, Michelle Peckham and Brendan Rogers)

Recent Highlights:

Myosins are important in regulating the organisation of the actin cytoskeleton:

myosins in cells

Our recent paper in Cell Reports (Makowska et al.,(2015) Cell Reports. 13, 2118-2125: link to article) shows that some myosins (myo1b, Myo9b, Myo10 and Myo18a) are overexpressed in metastatic prostate cancer. Specifically knocking down expression of each of these myosins result in the re-organisation of the actin cytoskeleton, and the type of reorganisation seen depends on which myosin has been depleted. Importantly, mis-regulation of myosin expression in these cells, helps to create the metastatic phenotype. (See also Michelle's recent review in Biochem. Soc. Trans (2016) on myosins in cancer http://www.biochemsoctrans.org/content/44/4/1026.long)

Super-resolution imaging of the Cilium

prgrip1L in the cilium

As part of a group working on the primary cilium, we have used dSTORM (direct Stochastic Optical Reconstruction Microscopy) on a home-built 3D set up at Leeds to image proteins in the transition zone. This example shows the protein RPGRIP1L. For more information see our recent paper Lambacher et al., (2016) TMEM107 recruits ciliopathy proteins to subdomains of the ciliary transition zone and causes Joubert syndrome. Nature Cell Biology, 18 p122-131 link to article

Live Cell Imaging: how does Myosin 10 get to the tip of filopodia?

myosin 10 in filopodium

111Mb, Quicktime 6Mb, Quicktime Myosin 10 is required to form filopodia, thin actin rich protrusions and it accumulates at the tip of these filopodia. It uses a combination of diffusion and active translocation to find its way to the right place in the cell. Tracking fluorescent myosin 10 molecules, and plotting their positions shows the tracks that this myosin uses in getting to the tip (as shown above). This image is taken from our recent paper in Journal of Biological Chemistry (Baboolal et al., (2016) J. Biol. Chem. 291, 22373-22385 link to article, and is a collaboration with Justin Molloy at the Crick Institute.

kymograph

watch this cool animation of myosin 10 moving to the tip, as an animated kymograph.

Improving muscle differentiation using specialised surfaces

myotubes

We've been working with the company 'Orla Protein Technologies' to improve myotube differentiation in culture and have found a combination of laminin 211 peptides, and FGF1 that gives good sarcomere striations (see above). See our recent paper (Parker et al., 2016, Cytotechnology, 68, pp2159-2169). We now have a PhD studentship available (BBSRC iCASE) to work with Orla on developing this technology further.

PhD projects for October 2017 start


older news


 

The Dynein homepage is now at www.dynein.org

 

 

111Mb, Quicktime 6Mb, Quicktime