Mass spectrometric differentiation of leucine and isoleucine in proteins derived from bacteria or cell culture

The differentiation of leucine and isoleucine is a well known difficulty in mass spectrometric peptide sequencing. A technique has been developed which allows these two amino acids to be distinguished by growing a bacterial or cell culture in a medium containing γ,δ-dl-dideuteroleucine. The isotopically labelled residue is incorporated into the cell's proteins, and the resulting mass spectra of leucine containing peptides exhibit sequence ions 2 amu higher than the corresponding isoleucine peptides.