Hepatic synthesis of carnitine from protein-bound trimethyl-lysine. Lysosomal digestion of methyl-lysine-labelled asialo-fetuin

Abstract

The biosynthesis of carnitine in the rat was studied by following the metabolism of two radioactive derivatives of asialo-fetuin. The first contained 14C-labelled methyl groups covalently bound to the 6-N-amino fraction of its lysine residues as 6-N-monomethyl- and dimethyl-lysine. By treating this protein with iodomethane, a second derivative was produced in which the radioactivity was preferentially incorporated as 6-N-[Me-14C]-trimethyl-lysine. These desialylated glycoproteins, like other asialo-proteins, were immediately cleared from the blood by rat liver. Within hepatocyte lysosomes, the 14C-labelled proteins were rapidly hydrolysed, producing free amino acids containing the various 6-N-[Me-14C]methylated lysine residues. The radioactive amino acids crossed the lysosomal membrane and were further metabolized in the cytosol. Carnitine was the major radioactive metabolite detected in extracts of the rat carcass and liver after intravenous injection of 6-N-[Me-14C]trimethyl-lysine-labelled asialo-fetuin. Within 3h, at least 34.6% of the trimethyl-lysine in the administered protein was converted into carnitine. Similarly, an isolated perfused rat liver converted 30% of the added peptide-bound trimethyl-lysine into carnitine within 90 min. On the other hand, in numerous attempts we failed to detect radioactive carnitine in both rat liver and carcass between 20 min and 22 h after injection of 6-N-[Me-14C]-monomethyl- and -dimethyl-lysine-labelled asialo-fetuin. These data provide evidence for a pathway of carnitine biosynthesis that involves trimethyl-lysine as a peptide-bound precursor as proposed by R.A. Cox & C.L. Hoppel [(1973) Biochem. J. 136, 1083-1090] and V. Tanphaichitr & H.P. Broquist [(1973) J. Biol. Chem. 248, 2176-2181]. The findings also show that rat liver can synthesize carnitine without the aid of other tissues, but cannot convert free partially methylated lysines into trimethyl-lysine.

Carnitine Supplementation in the Premature

“…Carnitine ultimately derives from the es sential amino acids lysine and methionine. The pathway of carnitine biosynthesis in mammals seems to be unique in that an obli gate intermediate, trimethyllysine is derived only from post-translational modification of peptide chains and subsequent breakdown of these proteins [48,49], However, in healthy volunteers oral administration of 5 g of ly sine was found to produce progressive rise in plasma trimethyllysine with an increase of plasma and urinary carnitine levels suggest ing the possibility of a non-peptidyl pathway of biosynthesis [50,51).…”

Variation of carnitine concentrations in Angus beef

“…Similarly, it has been shown that dietary carnitine enhances growth of rats fed a lowmethionine diet, indicating that dietary carnitine may possibly have a methionine-sparing effect (Khairallah and Wolf, 1965). On the other hand, protein that contains methylated lysine is required for carnitine synthesis (Labadie et al, 1976). Bremer (1961) showed that less of the methionine methyl groups is incorporated into carnitine.…”

Arsenic Speciation in Water and Human Urine by HPLC–ICP-MS and HPLC–MO–HG-AAS

“…The dry residue from urines were obtained through a cleanup procedure based on that of Labardie et al (15).…”