title: Expression and characterization of cathepsin P.
abstract: The mouse genome contains a family of clan C1A proteases that appear to be restricted to rodents within Eutherian (placental) mammals. mRNA analysis has shown that these genes are expressed exclusively in placenta. Sequence analysis predicts that the expressed proteins will be functional and consequently it was proposed that this family of proteases may have evolved to perform subspecialized functions of the closely related protease, cathepsin L, that is expressed in placental tissues of all mammalian species. In the present study, it was shown that cathepsin P can be expressed in Pichia pastoris as an inactive zymogen that can be activated with proteinase K, chymotrypsin or pancreatic elastase at neutral pH. Unlike other mammalian cathepsins, cathepsin P could also be autoactivated at neutral pH, but not at acidic pH. The activated enzyme was capable of hydrolysing peptidyl substrates and the protein substrates azocasein and transferrin, with optimal activity at pH 6.5-7.5. Little activity could be detected at pH 5.0 and below. Salts such as Na2SO4 and hyaluronate stimulated the activity of the protease against peptidyl substrates. The properties of cathepsin P appear to be quite distinct from those of cathepsin L, indicating that the duplication that gave rise to cathepsin P has probably not yielded an enzyme that provides a subfunction of cathepsin L in rodents. It seems probable that cathepsin P has evolved to perform a function that is performed by an evolutionarily unrelated protease in other mammalian species.
authors: Department of Biomedical Research, Alfred I. duPont Hospital for Children, 1600 Rockland Road, Wilmington, DE 19803, USA. mason@medsci.udel.edu
authors: Robert W
authors: RW
authors: Mason
authors: Carolyn A
authors: CA
authors: Bergman
authors: Guizhen
authors: G
authors: Lu
authors: Jennifer
authors: J
authors: Frenck Holbrook
authors: Katia
authors: K
authors: Sol-Church
mesh_terms: D000818:Animals; D056657:Cathepsin K; D002403:Cathepsins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D004792:Enzyme Precursors / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D051379:Mice; D010455:Peptides / Q000378:metabolism; D011480:Protease Inhibitors / Q000494:pharmacology; D011506:Proteins / Q000378:metabolism
chemical_list: D004792:Enzyme Precursors; D010455:Peptides; D011480:Protease Inhibitors; D011506:Proteins; D002403:Cathepsins; D056657:Cathepsin K; C534132:Ctsk protein, mouse