Figure 4.

DIAM identified myristoylglycine is synthesized during browning, being the only lapidated amino acid that induces brown adipocyte differentiation. (a–d) Dose–response effect of butyrylglycine and octanoylglycine (a), myristoyl alanine (b), oleoylphenylalanine, (c) and benzoylglycine and phenylacetylglycine (d) in brown adipocyte differentiation based on HCI data. (e) SGBS cells were labeled with 10 mM of [²H]-Myristic acid and [¹⁵N]-Glycine and allowed to differentiate with zafirlukast for 24 h. Afterward, metabolites were extracted and labeled and myristoylglycine was measured by LC/MS. De novo synthesis of myristoylglycine at the expense of labeled myristic acid was found during brown adipocyte differentiation by zafirlukast. (f) Human Glycine N-Acyltransferase (GLYAT) gene expression measured by qPCR at different time points. (g) Western blot analysis and quantification of human GLYAT in SGBS cells treated with zafirlukast at different time points.