Laeverin/aminopeptidase Q, a novel bestatin-sensitive leucine aminopeptidase belonging to the M1 family of aminopeptidases

J Biol Chem. 2007 Jul 13;282(28):20088-96. doi: 10.1074/jbc.M702650200. Epub 2007 May 24.

Abstract

Laeverin/aminopeptidase Q (APQ) is a cell surface protein specifically expressed on human embryo-derived extravillous trophoblasts that invades the uterus during placentation. The cDNA cloning of Laeverin/APQ revealed that the sequence encodes a protein with 990 amino acid residues, and Laeverin/APQ contains the HEXXHX(18)E gluzincin motif, which is characteristic of the M1 family of aminopeptidases, although the exopeptidase motif of the family, GAMEN, is uniquely substituted for the HAMEN sequence. In this study, we expressed a recombinant human Laeverin/APQ using a baculovirus expression system, purified to homogeneity, and characterized its enzymatic properties. It was found that Laeverin/APQ had a broad substrate specificity toward synthetic substrate, although it showed a preference for Leu-4-methylcoumaryl-7-amide. Searching natural substrates, we found that Laeverin/APQ was able to cleave the N-terminal amino acid of several peptides such as angiotensin III, kisspeptin-10, and endokinin C, which are abundantly expressed in the placenta. In contrast to the case with other M1 aminopeptidases, bestatin inhibited the aminopeptidase activity of Laeverin/APQ much more effectively than other known aminopeptidase inhibitors. These results indicate that Laeverin/APQ is a novel bestatin-sensitive leucine aminopeptidase and suggest that the enzyme plays important roles in human placentation by regulating biological activity of key peptides at the embryo-maternal interface.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs
  • Animals
  • Baculoviridae
  • CD13 Antigens / antagonists & inhibitors
  • CD13 Antigens / chemistry
  • CD13 Antigens / metabolism*
  • DNA, Complementary / genetics
  • DNA, Complementary / metabolism
  • Female
  • Gene Expression
  • Humans
  • Leucine / analogs & derivatives*
  • Leucine / pharmacology
  • Metalloproteases / chemistry
  • Metalloproteases / genetics
  • Metalloproteases / metabolism*
  • Placentation / physiology
  • Pregnancy
  • Protease Inhibitors / pharmacology*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism*
  • Substrate Specificity / drug effects
  • Trophoblasts / enzymology*

Substances

  • DNA, Complementary
  • Protease Inhibitors
  • Recombinant Proteins
  • Metalloproteases
  • laeverin, human
  • CD13 Antigens
  • Leucine
  • ubenimex