Sparse isotope labeling for nuclear magnetic resonance (NMR) of glycoproteins using 13C-glucose.

ABSTRACT: Preparation of samples for nuclear magnetic resonance (NMR) characterization of larger proteins requires enrichment with less abundant, NMR-active, isotopes such as 13C and 15N. This is routine for proteins that can be expressed in bacterial culture where low-cost isotopically enriched metabolic substrates can be used. However, it can be expensive for glycosylated proteins expressed in mammalian culture where more costly isotopically enriched amino acids are usually used. We describe a simple, relatively inexpensive procedure in which standard commercial media is supplemented with 13C-enriched glucose to achieve labeling of all glycans plus all alanines of the N-terminal domain of the highly glycosylated protein, CEACAM1. We demonstrate an ability to detect partially occupied N-glycan site

SUBMITTER: Rogals MJ 

PROVIDER: S-EPMC8091466 | biostudies-literature | 2021 May

REPOSITORIES: biostudies-literature