title: Partial purification and characterization of epidermal plasminogen activator and their inhibitor.
abstract: Plasminogen activator (PA) and PA inhibitor were partially purified from 2-d-old rat epidermis and characterized in order to elucidate the enzyme-inhibitor interaction in epidermis. PA extracted with buffer containing KSCN was first purified by Blue-Sepharose affinity chromatography. Separation of two PAs, with relative molecular mass (Mr) of 66000 and 44000, was accomplished by Con A-Sepharose column chromatography. The Mr 66000 enzyme had the properties of tissue-type PA (t-PA), while the Mr 44000 enzyme showed those of urokinase-type PA (u-PA) as determined by immunological and fibrin-binding studies. PA inhibitor was extracted in 1,4-piperazinediethanesulphonic acid buffer and purified by (NH4)2SO4 precipitation, gel filtration followed by a Mono Q column in an FPLC system. This inhibitor showed Mr 60000 and inhibited human u-PA activity in a dose- and time-dependent manner, but did not inhibit the activity of t-PA from human and murine melanoma cells or plasmin. It inhibited epidermal PA, Mr 44000, more effectively than it did the Mr 66000 epidermal PA. It was stable at 60 degrees C for 60 min or between pH 5 and 11. This study indicates that both u-PA and t-PA function in normal rat epidermis. On the other hand, an inhibitor which preferentially acts against u-PA exists, but inhibitor to t-PA does not appear to operate under normal epidermal functions.
authors.forename: M
authors.initials: M
authors.lastname: Nakagawa
authors.forename: K
authors.initials: K
authors.lastname: Fukuyama
authors.forename: W L
authors.initials: WL
authors.lastname: Epstein
authors.forename: A
authors.initials: A
authors.lastname: Hara
authors.forename: H
authors.initials: H
authors.lastname: Sawada
mesh_terms: D000818:Animals; D006801:Humans; D010960:Plasminogen Activators / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D015849:Plasminogen Inactivators; D051381:Rats; D012867:Skin / Q000378:metabolism*
chemical_list: D015849:Plasminogen Inactivators; D010960:Plasminogen Activators