The combined use of selective deuteration and double resonance experiments in assigning the 1H resonances of valine and tyrosine residues of dihydrofolate reductase

FEBS Letters

Roberts, G.C.K.¹  Searle, M.²  Birdsall, B.²  King, R.W.²  Griffiths, D.V.²  Feeney, J.²  Hammond, S.²  Kimber, B.² 

[1] Department of Chemistry, University ofKeele, Keele, Staffordshire ST5 5BG, England;Division of Physical Biochemistry, National Institute for Medical Research, Mill Hill, London UK. NW7 1AA

关键词: NMR;    Dihydrofolate reductase;    Deuterated protein;    NMR assignment;   

DOI  :  10.1016/0014-5793(84)80769-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Selective deuteration is a general solution to the resolution problem which limits the application of double resonance experiments to the assignment of the ¹H NMR spectra of proteins. Spin-decoupling and NOE experiments have been carried out on Lactobacillus casei dihydrofolate reductase and on selectively deuterated derivatives of the enzyme containing either [γ-²H₆]Val or (α,δ₂,ϵ₁-²H₃]His, [α,δ₁,δ₂,ϵ₁,ϵ₂,ζ-²H₆]Phe, [α,δ₁,ϵ₃,ζ₂,ζ₃,η₂-²H₆]Trp and [α,ϵ₁,ϵ₂-²H₃]Tyr. When combined with ring-current shift calculations based on the crystal structure of the enzyme, these experiments allow us to assign ¹H resonances of Val 61, Val 115, Tyr 46 and Tyr 68.

【 授权许可】

Unknown