De novo biosynthesis of enkephalins and their homologues in the human placenta

Fresh trophoblastic preparations of two human placentae delivered at term were pulse labelled for 30, 120 and 240 min with tritiated L-tyrosine. After deproteinizing and defatting, the peptide extracts were first concentrated through reversible hydrophobic binding on octadecasilyl-silica particles, prior to further resolution by repetetive high-performance liquid chromatography. Four peptides were isolated and purified to radioactive homogeneity, namely Met-enkephalin, Leu-enkephalin, (Arg⁶)-Leu-enkephalin, and (Arg⁶, Arg⁷)-Leu-enkephalin. Their presence and identity were further confirmed by substractive Edman degradation and by radioimmunoassay. No detectable amounts of radioactive Dynorphin could be trapped, however. Under the incubation conditions used, reference tritiated Leu-enkephalin had a biological half-life of circa 9.5 min.