Solid-state NMR structure determination of a membrane protein in E. coli cellular inner membrane.

ABSTRACT: Structure determination of membrane proteins in native cellular membranes is critical to precisely reveal their structures in physiological conditions. However, it remains challenging for solid-state nuclear magnetic resonance (ssNMR) due to the low sensitivity and high complexity of ssNMR spectra of cellular membranes. Here, we present the structure determination of aquaporin Z (AqpZ) by ssNMR in Escherichia coli inner membranes. To enhance the signal sensitivity of AqpZ, we optimized protein overexpression and removed outer membrane components. To suppress the interference of background proteins, we used a "dual-media" expression approach and antibiotic treatment. Using 1017 distance restraints obtained from two-dimensional ¹³C-¹³C spectra based on the comple

SUBMITTER: Xie H 

PROVIDER: S-EPMC10619923 | biostudies-literature | 2023 Nov

REPOSITORIES: biostudies-literature