Archives of Biochemistry and Biophysics 1997-08-01

A novel mammalian high-molecular-weight aminopeptidase.

H Erbeznik, L B Hersh

Index: Arch. Biochem. Biophys. 344(1) , 228-34, (1997)

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Abstract

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 approximately 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. Ap200 is relatively insensitive to inhibition by both puromycin, K(i) = 27 microM, and bestatin, K(i) = 1.6 microM. Among the synthetic beta-naphthylamides, Ap200 is more specific for alanine-beta-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 approximately 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.