Stereospecific assignments of the leucine methyl resonances in the 1H NMR spectrum of Lactobacillus casei dihydrofolate reductase
Open Access
- 1 March 1993
- journal article
- Published by Wiley in FEBS Letters
- Vol. 318 (2) , 177-180
- https://doi.org/10.1016/0014-5793(93)80016-n
Abstract
A general method is described for the stereospecific assignment of methyl resonances in protein NMR spectra based on selective deuteration procedures. A selectively deuterated dihydrofolate reductase from L. casei was prepared by incorporating stereoselectively deuterated l‐leucine, (2S,4R)[5,5,5‐2H3]leucine. By comparing the COSY spectra of the dihydrofolate reductase‐methotrexate complexes formed using deuterated and non‐deuterated enzyme the stereospecific assignments for resonances of all 13 leucine residues were obtained by noting the absence of cross‐peaks in spectra from the deuterated proteins.Keywords
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