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The combined use of selective deuteration and double resonance experiments in assigning the 1 H resonances of valine and tyrosine residues of dihydrofolate reductase
Author(s) -
Birdsall B.,
Feeney J.,
Griffiths D.V.,
Hammond S.,
Kimber B.,
King R.W.,
Roberts G.C.K.,
Searle M.
Publication year - 1984
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(84)80769-3
Subject(s) - valine , dihydrofolate reductase , tyrosine , resonance (particle physics) , chemistry , stereochemistry , enzyme , biochemistry , physics , amino acid , particle physics
Selective deuteration is a general solution to the resolution problem which limits the application of double resonance experiments to the assignment of the 1 H 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‐ 2 H 6 ]Val or (α,δ 2 ,ϵ 1 ‐ 2 H 3 ]His, [α,δ 1 ,δ 2 ,ϵ 1 ,ϵ 2 ,ζ‐ 2 H 6 ]Phe, [α,δ 1 ,ϵ 3 ,ζ 2 ,ζ 3 ,η 2 ‐ 2 H 6 ]Trp and [α,ϵ 1 ,ϵ 2 ‐ 2 H 3 ]Tyr. When combined with ring‐current shift calculations based on the crystal structure of the enzyme, these experiments allow us to assign 1 H resonances of Val 61, Val 115, Tyr 46 and Tyr 68.