A novel method of preparing totally alpha-deuterated amino acids for selective incorporation into proteins. Application to assignment of 1H resonances of valine residues in dihydrofolate reductase

FEBS Lett. 1990 Oct 15;272(1-2):197-9. doi: 10.1016/0014-5793(90)80483-y.

Abstract

The pyridoxal/2H2O exchange reaction of the alpha-CH of amino acids is known to be accompanied by racemisation: Thus by using a D-amino acid as the starting material any L-amino acid formed in the reaction will be essentially fully deuterated at its alpha-position. We have used this method to prepare alpha-deuterated L-valine and incorporated this biosynthetically into L. casei dihydrofolate reductase. A comparison of the alpha CH-NH fingerprint regions of COSY spectra of deuterated and normal DHFR complexes allows one to identify cross-peaks from 15 of the 16 valine residues.

MeSH terms

  • Amino Acids / metabolism*
  • Deuterium*
  • Isotope Labeling / methods*
  • Lacticaseibacillus casei / enzymology
  • Magnetic Resonance Spectroscopy*
  • Tetrahydrofolate Dehydrogenase / biosynthesis*
  • Tetrahydrofolate Dehydrogenase / chemistry
  • Valine / metabolism*

Substances

  • Amino Acids
  • Deuterium
  • Tetrahydrofolate Dehydrogenase
  • Valine