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

FEBS Lett. 1984 Oct 1;175(2):364-8. doi: 10.1016/0014-5793(84)80769-3.

Abstract

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 [gamma-2H6]Val or [alpha, delta 2, epsilon 1-2H3]His, [alpha, delta 1, delta 2, epsilon 1, epsilon 2, zeta-2H6]Phe, [alpha, delta 1, epsilon 3, zeta 2, zeta 3, eta 2-2H6]Trp and [alpha, epsilon 1, epsilon 2-2H3]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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Deuterium
  • Lacticaseibacillus casei / enzymology
  • Magnetic Resonance Spectroscopy / methods
  • Protein Binding
  • Protein Conformation
  • Tetrahydrofolate Dehydrogenase / metabolism*
  • Tyrosine / analysis*
  • Valine / analysis*

Substances

  • Tyrosine
  • Deuterium
  • Tetrahydrofolate Dehydrogenase
  • Valine