FEBS Letters | |
The combined use of selective deuteration and double resonance experiments in assigning the 1H resonances of valine and tyrosine residues of dihydrofolate reductase | |
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
【 预 览 】
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