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Stereospecific assignments of the leucine methyl resonances in the 1 H NMR spectrum of Lactobacillus casei dihydrofolate reductase
Author(s) -
Ostler G.,
Soteriou A.,
Moody C.M.,
Khan J.A.,
Birdsall B.,
Carr M.D.,
Young D.W.,
Feeney J.
Publication year - 1993
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(93)80016-n
Subject(s) - dihydrofolate reductase , lactobacillus casei , stereospecificity , deuterium , chemistry , leucine , stereochemistry , nmr spectra database , deuterium nmr , proton nmr , nuclear magnetic resonance spectroscopy , enzyme , spectral line , amino acid , biochemistry , physics , catalysis , quantum mechanics , astronomy , fermentation
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,4 R )[5,5,5‐ 2 H 3 ]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.