Folate status and age affect the accumulation of L-isoaspartyl residues in rat liver proteins

Formation of atypical L-isoaspartyl residues in proteins and peptides is a common, spontaneous and non-enzymatic modification of aspartyl and asparaginyl sites. The enzyme protein-L-isoaspartyl methyltransferase (PIMT) catalyzes the transfer of the methyl group of S-adenosyl-L-methionine (SAM) to these L-isoaspartyl sites, thereby allowing reisomerization and restoration of the original alpha peptide linkage. Because SAM is in part a product of folate metabolism, the present study was undertaken to determine the effects of folate deficiency on the presence of L-isoaspartyl residues in hepatic proteins. Young (weanling) and older (12 mo) Sprague-Dawley rats were fed a folate-sufficient (2 mg folate/kg diet) or folate-deficient (0 mg folate/kg diet) diet for 20 wk. Liver proteins were analyzed for L-isoaspartyl residues. This analysis was based on the PIMT-dependent incorporation of [[sup.3]H]-methyl groups from [[sup.3]H]-SAM and the subsequent (nonenzymatic) sublimation of these methyl groups into a nonaqueous scintillant. The amount of L-isoaspartyl residues in hepatic proteins was higher in younger folate-deficient than in folate-sufficient rats (deficient: 187[ + or -]71, sufficient: 64[+ or -]43 pmol/mg protein, P < 0.025). This difference, however, was not seen among the older groups of rats who instead exhibited a much larger accumulation of L-isoaspartyl residues in their hepatic proteins (deficient: 528[+ or -]151, sufficient: 470[+ or -]204 pmol/mg protein, P = 0.568). The importance of these observations is discussed. KEY WORDS: * folate * L-isoaspartyl * S-adenosyl-L-methionine * protein L-isoaspartyl methyltransferase * rats