Structure and Function of AAA ATPases

Project Overview

Example crystal structure of the AAA-domain of human Fidgetin-like 1 (PDB: 3D8B).

ATPases Associated with diverse cellular Activities (AAA) represent a diverse family of enzymes involved in a variety of fundamental cellular processes such as signal transduction and gene expression. AAA ATPases utilise the energy generated by nucleotide hydrolysis to remodel the structure of macromolecules or translocate proteins. Several AAA members are linked to the modulation of immunomodulatory processes and cancer progression. As such, these highly conserved proteins are of great interest to drug discovery and hence insight into their structure and function may contribute to the development of novel therapeutics.

The aim of the project is to express and purify of a small subfamily of AAA ATPases and obtain protein crystals for structure determination by X-ray crystallography. A further aspect of the proposed work is to study the function of these proteins by developing biochemical assays to support the development of small molecule tool compounds, so-called “chemical probes”, to enable the exploration of these proteins for drug discovery.

Training Opportunities

The successful candidate will be able to participate in a multidisciplinary project involving collaborations with academic partners as well as pharmaceutical companies. Taking advantage of the extensive experience in recombinant protein expression and X-ray crystallography at the Target Discovery Institute (TDI) and the Structural Genomics Consortium (SGC) Oxford, the successful candidate will be exposed to a variety of techniques such as high-throughput cloning, expression and purification of recombinant proteins in various host systems and protein crystallography. The project will rely on close interaction with different internal and external teams to pursue compound screens and further characterisation of this protein family by complementary cell biology approaches.

Theme

Protein Science & Structural Biology and Physiology, Cellular & Molecular Biology

Admissions

Project reference number: 780

Funding and admissions information

Supervisors

Name Department Institution Country Email
Dr Kilian Huber Structural Genomics Consortium Oxford University, NDM Research Building GBR kilian.huber@sgc.ox.ac.uk
Professor Paul Brennan Target Discovery Institute Oxford University, NDM Research Building GBR paul.brennan@sgc.ox.ac.uk

Hanson PI, Whiteheart SW. 2005. AAA+ proteins: have engine, will work. Nat. Rev. Mol. Cell Biol., 6 (7), pp. 519-29. Read abstract | Read more

The AAA+ (ATPases associated with various cellular activities) family is a large and functionally diverse group of enzymes that are able to induce conformational changes in a wide range of substrate proteins. The family's defining feature is a structurally conserved ATPase domain that assembles into oligomeric rings and undergoes conformational changes during cycles of nucleotide binding and hydrolysis. Here, we review the structural organization of AAA+ proteins, the conformational changes they undergo, the range of different reactions they catalyse, and the diseases associated with their dysfunction. Hide abstract

Arrowsmith CH, Audia JE, Austin C, Baell J, Bennett J, Blagg J, Bountra C, Brennan PE, Brown PJ, Bunnage ME, Buser-Doepner C, Campbell RM, Carter AJ, Cohen P, Copeland RA, Cravatt B, Dahlin JL, Dhanak D, Edwards AM, Frederiksen M, Frye SV, Gray N, Grimshaw CE, Hepworth D, Howe T, Huber KV, Jin J, Knapp S, Kotz JD, Kruger RG, Lowe D, Mader MM, Marsden B, Mueller-Fahrnow A, Müller S, O'Hagan RC, Overington JP, Owen DR, Rosenberg SH, Roth B, Ross R, Schapira M, Schreiber SL, Shoichet B, Sundström M, Superti-Furga G, Taunton J, Toledo-Sherman L, Walpole C, Walters MA, Willson TM, Workman P, Young RN, Zuercher WJ. 2015. The promise and peril of chemical probes. Nat. Chem. Biol., 11 (8), pp. 536-41. | Read more