NMR Resonance Assignment Methodology: Characterizing Large Sparsely Labeled Glycoproteins.

ABSTRACT: Characterization of proteins using NMR methods begins with assignment of resonances to specific residues. This is usually accomplished using sequential connectivities between nuclear pairs in proteins uniformly labeled with NMR active isotopes. This becomes impractical for larger proteins, and especially for proteins that are best expressed in mammalian cells, including glycoproteins. Here an alternate protocol for the assignment of NMR resonances of sparsely labeled proteins, namely, the ones labeled with a single amino acid type, or a limited subset of types, isotopically enriched with ¹⁵N or ¹³C, is described. The protocol is based on comparison of data collected using extensions of simple two-dimensional NMR experiments (correlated chemical shifts, nuclear Overhau

SUBMITTER: Chalmers GR 

PROVIDER: S-EPMC6554063 | biostudies-literature | 2019 May

REPOSITORIES: biostudies-literature