Application of phorbol ester to mouse skin causes a rapid and sustained loss of protein kinase C

Abstract

It is now widely accepted that tumour-promoting phorbol esters activate a Ca²⁺- and phospholipid-dependent protein kinase (protein kinase C) both in vitro and in intact cells^1–3, and that the kinase represents a major cellular phorbol ester-binding protein. The phorbol esters act as analogues of diacylglycerol, a natural regulator of protein kinase C, and stabilize the membrane-association of the kinase. Although other molecular targets may exist, protein kinase C activation is probably important in mediating the diverse responses of cultured cells to phorbol esters and in promoting in vivo tumours. The enzyme comprises a family of closely related proteins^4–7 and has been detected in extracts from mouse epidermal cells^8–10, the likely targets for two-stage car-cinogenesis in mouse skin. In this report we show that application of a single dose of TPA (12-O-tetradecanoyl phorbol-13-acetate) to mouse skin results in a rapid and complete loss of protein kinase C activity which is maintained for 3–4 days. This is associated with a loss of immunologically detectable protein kinase C and the accumulation of a smaller protein detectable by antibodies recognizing the regulatory domain of protein kinase C.