Synthesis of [u-13C,15N]-cysteine hydrochloride: An important tool for heteronuclear, multi-dimensional NMR studies of proteins
✍ Scribed by Michael J. Panigot; Stephen W. Fesik; Robert W. Curley Jr.
- Book ID
- 102371199
- Publisher
- John Wiley and Sons
- Year
- 1995
- Tongue
- French
- Weight
- 310 KB
- Volume
- 36
- Category
- Article
- ISSN
- 0022-2135
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✦ Synopsis
Protein structure determination by modern NMR techniques is greatly facilitated using 15N-and "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 ~ells,[U-'~C,'~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-"C,'5N]-cysteine hydrochloride in good overall yield is described which makes use of readily available "N-and "C-labeled starting materials and will facilitate heteronuclear multidimensional NMR studies of proteins that are overexpressed in mammalian cells.