Methionine-dependence phenotype of tumors:: Metabolite profiling in a melanoma model using L-[methyl-13C]methionine and high-resolution magic angle spinning 1H-13C nuclear magnetic resonance spectroscopy

Tumors frequently have abnormal L-methionine (Met) metabolism, the so-called Met-dependence phenotype that refers to the inability to proliferate in the absence of Met. However, the origin of this phenotype is still unknown and may arise from one of several pathways of Met metabolism. To help characterize the metabolic features of Met-dependent/independent phenotypes, the fate of the methyl carbon of L-[methyl-C-13]Met was chased in a murine model of malignant melanoma (B16-F1) in vitro and in vivo. Growth curves under Met restriction showed that melanoma cells in vitro were Met-independent, whereas implanted melanoma tumors in vivo were Met-dependent. Label-assisted high-resolution magic angle spinning H-1-C-13 NMR spectroscopy metabolite profiling showed that, in vitro, creatine and phosphatidylcholine C-13-enrichments were poor, but S-adenosyl-Met and posttranslationally N-methylated protein signals were strong. In contrast, in vivo, creatine and phosphatidylcholine enrichments were strong but S-adenosyl-Met and N-methylated protein signals were poor. In addition, in vivo, transsulfuration was very efficient, consumed one-carbon units originating from the methyl carbon of Met, and yielded taurine labeling. From these data, the Met-dependent/independent phenotypes appear closely related to the source of one-carbon units. Thus, L-[methyl-C-13]Met-assisted NMR spectroscopy metabolite profiling allowed the discrimination between Met-dependence and Met-independence and provided novel mechanistic information on their origin.