title: 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.
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.
authors.affiliation: Laboratory of Molecular Structure, National Institute for Medical Research, London, UK.
authors.forename: J
authors.initials: J
authors.lastname: Feeney
authors.forename: B
authors.initials: B
authors.lastname: Birdsall
authors.forename: G
authors.initials: G
authors.lastname: Ostler
authors.forename: M D
authors.initials: MD
authors.lastname: Carr
authors.forename: M
authors.initials: M
authors.lastname: Kairi
mesh_terms: D000596:Amino Acids / Q000378:metabolism*; D003903:Deuterium; D007553:Isotope Labeling / Q000379:methods*; D007780:Lacticaseibacillus casei / Q000201:enzymology; D009682:Magnetic Resonance Spectroscopy; D013762:Tetrahydrofolate Dehydrogenase / Q000096:biosynthesis* / Q000737:chemistry; D014633:Valine / Q000378:metabolism*
chemical_list: D000596:Amino Acids; D003903:Deuterium; D013762:Tetrahydrofolate Dehydrogenase; D014633:Valine