In-cell monitoring of membrane proteins at atomic-resolution


Protein function depends on environment. The ultimate goal of structure biology is hence to understand protein function in its native environment, which is the cell. This is particularly true for membrane proteins. Cellular membranes are complex (> 1000 lipid species), interspersed with nanodomains and crowded with proteins. This overwhelming intricacy, difficult to define and impossible to replicate in vitro, is critical for membrane protein function. But because of the enormous challenge to resolve proteins in such heterogeneous environments, structure biology hitherto lacks approaches to study proteins in cellular membranes and principal questions on the physiological states of membrane proteins remain unanswered.
Here, I propose to develop a methodological ensemble of solid-state NMR and computational microscopy to monitor proteins in cellular membranes at atomic detail. I will capitalize on my recent developments and strong expertise in ultra-sensitive proton-detected solid-state NMR to selectively resolve proteins in crowded native membranes. Complemented by large- and multi-scale molecular dynamics simulations, my project will provide an atomic-level perspective on supramolecular structures and dynamics of proteins in cellular membranes. My project will lead to a major step forward in structure biology, since the capacity to observe native membrane proteins at work holds the keys to truly understanding biological processes and paves the way for powerful applications such as high-resolution drug-binding assays in cellular membranes.
My approach will be implemented on a small lipoprotein. In comparative cellular and in vitro studies, the tested methods will then be used to investigate the structure-dynamics-function relationship in pharmaceutically important membrane transporters with a focus on the influence of the native lipid nanoenvironment on transport, which is completely unknown. The limits of cellular solid-state NMR will be explored with a large secretion system, that is found in many pathogenic bacteria and that has resisted structure elucidation for the past 15 years.


Wetenschappelijk artikel

Hoofdstuk in boek

  • D Mance, S Narasimhan, C Pinto, M Baldus, R Ghose, AMJJ Bonvin, M Weingarth(2018): Protein NMR pp. 111 - 132





Dr. M. Weingarth

Verbonden aan

Universiteit Utrecht, Faculteit Bètawetenschappen, Departement Scheikunde


J. Medeiros Silva MSc


01/11/2015 tot 31/10/2020