Research
Summary
The cytoplasmic dynein complex, along with its cofactor dynactin and other regulatory proteins, acts as a motor that moves along microtubules. Among its many functions, dynein organises membraneous compartments, gathers up misfolded proteins, carries viruses and plays key roles in cell division and replication.
Despite playing such a prominent and crucial role in numerous biological and disease processes, very little structural information is known for cytoplasmic dynein. Challenges such as its large size, complexity and low abundance mean that it represents one of the major unexplored and exciting frontiers in structural biology.
The goal of our group is to use a combination of X-ray crystallography, protein engineering and advanced single molecule techniques to further our understanding of the structure and mechanism of the dynein mega complex. This will allow us to answer questions such as how it moves along microtubules, how it is regulated and how it interacts with cargos. The long-term hope is that this information will aid the development of therapeutics that will block certain dynein functions (such as transport of viruses) without inhibiting the whole motor.
Please feel free to follow the links below for more information about dynein and highlights of Andrew’s previous research. Alternatively the whole section can be downloaded as a pdf .
Contents
- Background
- The dynein family (cytoplasmic and axonemal dyneins)
- Cytoplasmic dynein roles in the cell and disease
- Components of the cytoplasmic dynein complex
- The dynein motor domain
- Research Highlights
- Recombinant expression of dynein
A significant hurdle to studying the dynein motor has been its sheer size. One of the major breakthroughs in recent years has been the ability to recombinantly express and manipulate the dynein gene. This section describes the use of homologous recombination in Saccharomyces cerevisiae to generate an artificially dimerised minimal dynein motor. This work forms the basis for many biochemical, biophysical and protein engineering experiments.
- Single molecule studies
It is now possible to determine the position of a fluorophore with nanometer precision. At this scale it is possible to get direct information about the movements within proteins using light microscopy. My colleagues and I were able to use these methodologies to follow the stepping of dynein as it moves along microtubules.
- Structure of the dynein stalk
We have recently reported the crystal structure of the dynein microtubule binding domain. This is the first part of the dynein motor to be solved and gives insight into long range communication within the dynein molecule.
- The dynein family (cytoplasmic and axonemal dyneins)
- Cytoplasmic dynein roles in the cell and disease
- Components of the cytoplasmic dynein complex
- The dynein motor domain
- Recombinant expression of dynein
- Single molecule studies
- Structure of the dynein stalk
A significant hurdle to studying the dynein motor has been its sheer size. One of the major breakthroughs in recent years has been the ability to recombinantly express and manipulate the dynein gene. This section describes the use of homologous recombination in Saccharomyces cerevisiae to generate an artificially dimerised minimal dynein motor. This work forms the basis for many biochemical, biophysical and protein engineering experiments.
It is now possible to determine the position of a fluorophore with nanometer precision. At this scale it is possible to get direct information about the movements within proteins using light microscopy. My colleagues and I were able to use these methodologies to follow the stepping of dynein as it moves along microtubules.
We have recently reported the crystal structure of the dynein microtubule binding domain. This is the first part of the dynein motor to be solved and gives insight into long range communication within the dynein molecule.
