N6-Trimethyl-lysine metabolism. Structural identification of the metabolite 3-hydroxy-N6-trimethyl-lysine.

Abstract

(1)H and (13)C nuclear-magnetic-resonance spectroscopy and functional-group analysis were used to determine the molecular structure of an isolated metabolite (II(b)) of trimethyl-lysine as 3-hydroxy-N(6)-trimethyl-lysine, an important intermediate in the conversion of trimethyl-lysine into trimethylammoniobutyrate and carnitine [Hoppel, Cox & Novak (1980) Biochem. J.188, 509-519]. Functional-group analysis revealed the presence of a primary amine and reaction of metabolite (II(b)) with periodate yielded 4-N-trimethylammoniobutyrate as a product, showing 2,3-substitution on the molecule and suggesting that the 3-substitution on the molecule may be an alcohol ([unk]CH-OH), amine ([unk]CH[unk]-NH(2)) or carbonyl ([unk]C=O) functional group. (1)H integration ratios, (1)H and (13)C chemical-shift data and (1)H and (13)C signal multiplicities from the sample (II(b)) were used to complete the identification of metabolite (II(b)) as 3-hydroxy-N(6)-trimethyl-lysine. For example, the proton multiplet at delta 4.2p.p.m. and doublet at delta 4.1p.p.m., positions representative of amine or alcohol substitution on methylene carbon atoms, integration ratios of 1:1:2:9:4 and a positive ninhydrin test suggest 3-hydroxy-N(6)-trimethyl-lysine as the molecular structure for metabolite (II(b)). (13)C chemical-shift data obtained from the sample (II(b)) and compared with several model compounds (trimethylammoniohexanoate, trimethyl-lysine and 3-hydroxylysine) resulted in generation of the spectrum of the metabolite and allowed independent identification of metabolite (II(b)) as 3-hydroxy-N(6)-trimethyl-lysine. The (1)H spectrum of erythro- and threo-3-hydroxylysine are presented for comparison, and the (1)H and (13)C n.m.r. spectra of the erythro-isomer support this analysis.