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Synthesis of [ u ‐ 13 C, 15 N]‐cysteine hydrochloride: An important tool for heteronuclear, multi‐dimensional NMR studies of proteins
Author(s) -
Panigot Michael J.,
Fesik Stephen W.,
Curley Robert W.
Publication year - 1995
Publication title -
journal of labelled compounds and radiopharmaceuticals
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.432
H-Index - 47
eISSN - 1099-1344
pISSN - 0362-4803
DOI - 10.1002/jlcr.2580360506
Subject(s) - chemistry , heteronuclear molecule , cysteine , amino acid , hydrochloride , carbon 13 nmr , nuclear magnetic resonance spectroscopy , stereochemistry , biochemistry , enzyme
Protein structure determination by modern NMR techniques is greatly facilitated using 15 N‐ and 13 C‐labeled proteins. Labeling of proteins that are overexpressed in mammalian cells is a difficult task that requires a growth medium consisting of algal hydrolysates supplemented with labeled amino acids. Although most of the amino acids can be obtained to prepare an isotopically labeled growth medium for mammalian cells,[ U ‐ 13 C, 15 N]‐cysteine is not available, hampering the backbone and cysteine side‐chain assignments and structure determination in the vicinity of the cysteine residues. A synthesis of D,L‐[ U ‐ 13 C, 15 N]‐cysteine hydrochloride in good overall yield is described which makes use of readily available 15 N‐ and 13 C‐labeled starting materials and will facilitate heteronuclear multidimensional NMR studies of proteins that are overexpressed in mammalian cells.