In vitro degradation of oxytocin by pregnancy serum, placental subcellular fractions and purified placental aminopeptidases.
The degradation of oxytocin (formula; see text) 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 placental 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 Ile3 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.(ABSTRACT TRUNCATED AT 250 WORDS)
D000596:Amino Acids / Q000032:analysis; D000626:Aminopeptidases / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D010450:Endopeptidases / Q000378:metabolism; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D007931:Leucyl Aminopeptidase / Q000378:metabolism; D008861:Microsomes / Q000201:enzymology; D010121:Oxytocin / Q000097:blood / Q000378:metabolism*; D010920:Placenta / Q000201:enzymology / Q000378:metabolism*; D011247:Pregnancy; D013347:Subcellular Fractions / Q000201:enzymology*
D000596:Amino Acids; D010121:Oxytocin; D010450:Endopeptidases; D000626:Aminopeptidases; D007931:Leucyl Aminopeptidase