Title of project: Combating Inheritable heart disease: Functional and Molecular
characterisation of Mitochondrial ADP/ATP carrier
Director of Studies: Dr Vincent Postis
Second Supervisor: Prof Guy Lauquin
Overview of project
Mitochondrial adenine nucleotide carriers are polytopic proteins located in the inner mitochondrial
membrane. They are essential for life as they are one of the main sources of ATP supply for the cell
thanks to their ability to export matrix ATP, synthesised by the respiratory chain, and import cytosolic
ADP, thus providing new substrate for the ATP synthase. Three tissue specific isoforms exist in
humans. Point mutations in the human cardiac isoform (hANC1) have been shown to be involved in
two main pathologies: myopathies and autosomal dominant progressive external ophtalmoplegia
(adPEO). The first affect muscles and the second is clinically characterized by exercise intolerance,
ptosis and muscle weakness. The mechanisms underlying the mitochondrial pathologies caused by
hANC1 mutations remain largely poorly understood.
In the light of extensive biochemical studies, it is admitted that these transporters switch between at
least four conformations (ADP binding/release and ATP binding/release), two of which can be frozen
by the use of specific inhibitors called carboxyatractylate (CATR) and bongkrekic acid (BA). In 2003,
the molecular structure of the bovine homologue of hAnc1 has been solved in a CATR conformation,
allowing to understand one of the conformations the transporter adopt during the ADP/ATP exchange.
The complete cycle, however, has not been yet understood due in part to the inability to isolate a BAinhibited conformation. In addition, although more than 20 years of biochemical data tend to indicate
that these carriers function as dimers, the crystal structure only showed a monomer and opened an
endless debate.
The aim of the project is to isolate and structurally characterise the adenine nucleotide translocators
through the solving of their structure in a BA conformation as well as isolating the transporter in his
native lipidic environment. Unlike conventional techniques using detergents which have failed to
produce crystal of this transporter in its 'BA conformation', this study will use a cutting edge
methodology which take advantage of the SMALP (styrene maleimic acid lipoparticles) technique.
To maximise the applicant chance of success, the human ANC1 protein will be studied together with
the yeast homologue ANC2. Both isoforms genes are readily available from Dr Postis previous work
(de Marcos Lousa et al., 2003; Postis et al., 2004). These will need to be cloned in an adequate
vector for expression in mammalian cells such as HEK cells, which have been demonstrated to be a
reliable system for membrane protein expression. Molecular studies using SMALP technology has
been pioneered by Dr Postis (Postis et al., 2015). Hence this project will combine basic molecular
biology and biochemistry, ideal techniques to be learned by a PhD student for future employability.
Link to Faculty Research Themes
School of Rehabilitation and Health Sciences 5A (Biological Sciences)
Outline of project including proposed timescales
This project will give the opportunity to the PhD student's to develop skills in a cutting edge
methodology highly looked at by the pharmaceutical industry. Finally, the structure resolution will be
solved in collaboration with prof. Goldman, which is a renowned scientist in the crystallography field.
This will allow the PhD student to create a network for collaborators for future job/grant applications.
This work outcome will be disseminated through publication in high impact journals, such as JBC
(impact factor 4.6) and JMB (impact factor 3.9). The results obtained during the first year of this
program will be used as support for a BBSRC New Investigator Scheme application for Dr Postis
Further information
To apply you must be eligible for NHS Continuing Professional Development (CPD) funding and have
the support of your line manager in writing. General enquiries should be directed by email to the
Faculty Research Director r.hogston@leedsbeckett.ac.uk to discuss the project further please contact
the Director of Studies V.L.Postis@leedsbeckett.ac.uk
Applications should be made on line here
http://www.leedsbeckett.ac.uk/research/research-degrees/research-studentships-andfees-only-bursaries/