Stereospecific assignments of the leucine methyl resonances in the 1H NMR spectrum of Lactobacillus casei dihydrofolate reductase

FEBS Letters

Ostler, G.¹  Carr, M.D.¹  Khan, J.A.²  Young, D.W.²  Soteriou, A.¹  Birdsall, B.¹  Feeney, J.¹  Moody, C.M.² 

[1] Laboratory of Molecular Structure, National Institute for Medical Research, Mill Hill, London NW7 1AA, UK;School of Chemistry and Molecular Sciences, The University of Sussex, Falmer, Brighton BN1 9QJ, UK

关键词: Dihydrofolate reductase;    Stereospecific NMR assignment;    Deuterated leucine;   

DOI  :  10.1016/0014-5793(93)80016-N

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown