Synthesis of ¹³C-methyl-labeled amino acids and their incorporation into proteins in mammalian cells.

ABSTRACT: Isotopic labeling of methyl-substituted proteinogenic amino acids with ¹³C has transformed applications of solution-based NMR spectroscopy and allowed the study of much larger and more complex proteins than previously possible with ¹⁵N labeling. Procedures are well-established for producing methyl-labeled proteins expressed in bacteria, with efficient incorporation of ¹³C-methyl labeled metabolic precursors to enable the isotopic labeling of Ile, Val, and Leu methyl groups. Recently, similar methodology has been applied to enable ¹³C-methyl labeling of Ile, Val, and Leu in yeast, extending the approach to proteins that do not readily fold when produced in bacteria. Mammalian or insect cells are nonetheless preferable for production of many human

SUBMITTER: Borgini M 

PROVIDER: S-EPMC10825848 | biostudies-literature | 2023 Nov

REPOSITORIES: biostudies-literature