A significant challenge for the use of liquid state NMR spectroscopy for the structure determination of membrane proteins is sample preparation of the membrane proteins in environments that are suitable for investigation by solution NMR and keep the protein in a functional state. Detergents have long been the mainstay of researchers but often to the detriment of protein function. Recent work has seen an expansion in the number of membrane mimetics such as the nanodiscs or bicelles. The large size of nanodiscs, however, hinders their broad application in solution NMR characterization of membrane proteins with integral membrane proteins resulting in broad and overlapping signals. We have developed a detergent titration method that significantly enhances the spectral quality combined with a combinatorial labeling strategy that facilitates the backbone assignment of the integral membrane protein lipoprotein signal peptidase II. Our method thereby facilitates NMR investigations while preserving the central benefit of nanodisc technology i.e. the presence of a lipid core around the MP necessary for function.<br/>

Reverse q-titration of integral membrane proteins in nanodiscs

254th National Meeting (2017)

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