Mass photometry enabled label-free platform to study the dynamics of membrane proteins assembly

Aston Institute for Membrane Excellence

At Aston Institute for Membrane Excellence (AIME), we are passionate about equity, diversity, and inclusion and are dedicated to cultivating a diverse cohort of PhD researchers from a range of academic backgrounds.

AIME is a first-of-its-kind institute that synergistically combines world-leading expertise in membrane biology and novel polymer chemistries. No other institute in the global research and innovation landscape has the platform, potential or promise for major breakthroughs that AIME heralds.

We welcome students from diverse pathways, whether recent graduates or professionals returning to academia.

We are particularly focussed on recruiting and training female scientists in this exciting multi-disciplinary arena, acknowledging current gender disparities. All AIME PhD projects feature:

• Flexible working.
• Comprehensive mentorship for all students.
• Supported opportunities for secondments in international labs.
• Diverse scientific and transferrable skills training, equipping all graduates for leading roles in academia or industry.

 If you have any questions about this opportunity, please contact p.r.knobbs@aston.ac.uk, AIME Manager for a chat.

Details of the Project

Integral membrane proteins (IMPs) are the gatekeepers to cell function and their importance is outlined by their significant role in variety of diseases including viral infections, various physiological diseases such as diabetes, Alzheimer’s, Parkinson’s, Cystic Fibrosis, and a range of cancers. IMPs collectively represents more than 50% of existing drug targets, however, studying them poses significant challenges. IMPs are inserted in lipid bilayers, and both bilayer composition and thickness govern IMP oligomeric nature, conformational changes and protein interactions. However, IMP characterisation through existing techniques relies on extracting IMPs from their native environment, thus removing the physiological context resulting in IMP instability, misfolding, loss of oligomerisation and protein interactions. 

To overcome these challenges, this project will employ interdisciplinary approach to combine polymer synthesis, protein expression and purification, microfluidics, membrane-biochemistry and single molecule imaging to develop a platform that can study IMPs in native bilayer environment and provides a rapid assay to screen drugs and antibodies therapies against IMPs.

The project is divided into two-steps, in the first step, polymer supported bilayers (cushioned bilayers) will be developed. This step will combine polymer chemistry with membrane biochemistry and cushioned bilayers will be extensively characterised using fluorescence microscopy and single-particle-tracking with mass photometry (Dynamic MP).

The second step of the project will involve development of protocol to insert membrane proteins into cushioned bilayers. This project will focus on inserting aquaporins, GPCRs and ion channels into the cushioned bilayers. For each protein, the insertion mechanism and oligomerisation dynamics will be studied using Dynamic MP. This step will provide the molecular understanding of IMP characteristics including oligomerisation and oligomer specific lateral diffusion in the bilayer.

The oligomerisation and diffusion correlation will establish an IMP specific characteristic that illustrates the effect of lipid environment on IMP protein-protein interaction and conformational changes. Furthermore, oligomer to diffusion correlation is a unique aspect of the platform that is out of reach for existing methodologies and integration of this parameter with existing IMP research will provide a wholistic perspective of IMP structure, interaction and function.

To validate the relevance of the platform for drug screening, IMP specific small molecules and monovalent antibodies will be used to estimate their respective affinity and effect on IMP oligomer dynamics and lateral diffusion.

Development of the platform will have significant impact on IMP research and will establish a basis to study wide variety of IMPs in their native bilayer environment.

The PhD project provides an excellent opportunity to develop both experimental and analytical expertise. The candidate will be supported by the interdisciplinary team at AIME and will be working with the leading edge of technologies including polymer synthesis, membrane protein research and single-particle-tracking using mass photometry at AIME. Candidates will be provided with appropriate training and support to carry out the project and gain skills to succeed career in academics or in industry.

At AIME, we strive to create an environment that fosters the growth and development of candidates from diverse backgrounds and encourage individuals of all genders to apply.

Person Specification

The successful applicant should have been awarded, or expect to achieve, a Masters degree in a relevant subject with a 60% or higher weighted average, and/or a First or Upper Second Class Honours degree (or an equivalent qualification from an overseas institution) in a relevant subject. Good theoretical knowledge of biochemistry, microscopy, enzyme kinetics, lipid bilayers. Practical experience with microbiology and protein expression and purification is desirable but not necessary. Knowledge of coding would be beneficial. 

Contact information

For formal enquiries about this project contact Manish Singh Kushwah at m.kushwah@aston.ac.uk

Submitting an application

 We can only consider applications that are complete and have all supporting documents. Applications that do not provide all the relevant documents will be automatically rejected. Your application must include:

1. English language copies of the transcripts and certificates for all your higher education degrees, including any Bachelor degrees.
2. A Research Statement detailing your understanding of the research area, how you would approach the project, and a brief review of relevant literature. Be sure to use the title of the research project you are applying for. There is no set format or word count.
3. A personal statement which outlines any further information which you think is relevant to your application, such as your personal suitability for research, career aspirations, possible future research interests, and further description of relevant employment experience.
4. Two academic referees who can discuss your suitability for independent research. References must be on headed paper, signed and dated no more than 2 years old. At least one reference should be from your most recent University. You can submit your references at a later date if necessary.
5. Evidence that you meet the English Language requirements. If you do not currently meet the language requirements, you can submit this at a later stage.
6. A copy of your passport. Where relevant, include evidence of settled or pre-settled status.

Apply for this position here

If you require further information about the application process, please contact the Postgraduate Admissions team at pgr_admissions@aston.ac.uk