A Novel Mammalian High-Molecular-Weight Aminopeptidase

Studies with the human lymphoma U937 cell line revealed the presence of two soluble aminopeptidase activities. Using specific antisera one of these was identified as the puromycin-specific aminopeptidase, while the other appeared to be a novel ∼200-kDa activity. The kinetic properties of this high-molecular-weight aminopeptidase, referred to as Ap200, were similar to those of the puromycin-sensitive aminopeptidase, but showed quantitative differences. Ap200is relatively insensitive to inhibition by both puromycin,Ki= 27 μm, and bestatin,Ki= 1.6 μm. Among the synthetic β-naphthylamides, Ap200is more specific for alanine-β-naphthylamide compared to the puromycin-sensitive aminopeptidase. Similarly, this enzyme cleaves a more limited number of physiological peptides exhibiting a preference for the enkephalins. Ammonium sulfate, but not sodium chloride at the same ionic strength, was able to dissociate the high-molecular-weight aminopeptidase to a ∼100-kDa active form. The high-molecular-weight aminopeptidase is found as a low abundant protein in a number of tissues including intestine, kidney, liver, lung, muscle, spleen, and testes, but could not be detected in adrenal, heart, or brain. Thus, it has a tissue distribution which differs from the puromycin-sensitive aminopeptidase.