In Vitro Degradation of Oxytocin by Pregnancy Serum, Placental Subcellular Fractions and Purified Placental Aminopeptidases

Summary

The degradation of oxytocin (Cys¹-Tyr²-Ile³-Gln⁴-Asn⁵-Cys⁶-Pro⁷-Leu⁸-Gly⁹-NH₂)

by human placental particulate and soluble fractions, pregnant and non-pregnant sera, and purified enzymes such as microsomal placental leucine aminopeptidase (P-LAP), retroplacental serum P-LAP and placental post-proline endopeptidase, was studied by measuring liberated amino acids with a high performance liquid Chromatograph (HPLC). While the placental particulate fraction degraded oxytocin almost completely into single amino acid and amide, the placental soluble fraction did not liberate any amino acid and amide. Purified retroplacental P-LAP liberated Tyr², Ile³, Gln⁴ and Asn⁵ from the cyclic structure of oxytocin actively. Pregnancy serum containing the retroplacental P-LAP liberated also these amino acids and amides. Purified microsomal P-LAP liberated Leu⁸ in addition to these amino acids and amides. Although purified placental post-proline endopeptidase or porcine kidney leucine aminopeptidase (LAP) could not liberate any amino acid and amide from oxytocin, the combination of the post-proline endopeptidase with porcine kidney LAP or with pla-cental microsomal P-LAP actively liberated all amino acids and amides detectable by HPLC. When the ratio of amino acid liberation velocity to LAP activity measured with leu-p-nitroanilide was calculated, the ratios for cyclic amino acids such as Tyr² and He³ were high with the placental particulate fraction, the mixture of post-proline endopeptidase and microsomal P-LAP, retroplacental P-LAP, and pregnancy serum. The ratio for Leu⁸ was high with the placental particulate fraction and the mixture of post-proline endopeptidase and microsomal P-LAP. It was concluded that the microsomal P-LAP plus placental post-proline endopeptidase seem a great contribution to the degradation of oxytocin in human placenta and its degradation in pregnancy serum is due to the retroplacental P-LAP.

Key Words

Oxytocin - microsomal placental leucine aminopeptidase (P-LAP) - retroplacental P-LAP - placental post-proline endopeptidase - high performance liquid chromatography