Sortase-mediated segmental labeling: A method for segmental assignment of intrinsically disordered regions in proteins.

ABSTRACT: A significant number of proteins possess sizable intrinsically disordered regions (IDRs). Due to the dynamic nature of IDRs, NMR spectroscopy is often the tool of choice for characterizing these segments. However, the application of NMR to IDRs is often hindered by their instability, spectral overlap and resonance assignment difficulties. Notably, these challenges increase considerably with the size of the IDR. In response to these issues, here we report the use of sortase-mediated ligation (SML) for segmental isotopic labeling of IDR-containing samples. Specifically, we have developed a ligation strategy involving a key segment of the large IDR and adjacent folded headpiece domain comprising the C-terminus of A. thaliana villin 4 (AtVLN4). This procedure significantly reduces the complexi

SUBMITTER: Boyko KV 

PROVIDER: S-EPMC8553144 | biostudies-literature | 2021

REPOSITORIES: biostudies-literature