Phorbol ester, calcium ionophore, or serum added to quiescent rat embryo fibroblast cells all result in the elevated phosphorylation of two 28,000-dalton mammalian stress proteins.

Rat embryo fibroblast cells grown under stress (e.g. heat shock, arsenite, or amino acid analogue treatment) show elevated levels of a number of proteins with apparent molecular masses between 28,000-110,000 daltons (i.e. stress proteins). It is shown that the smaller 28,000-dalton stress proteins, which do not contain methionine, are comprised of at least four isoforms, all of which appear related as determined by one-dimensional peptide mapping. 32PH3PO4 labeling of normal and stressed cells demonstrates that three of the four 28-kDa isoforms are phosphoproteins. In the course of other studies phosphorylation of two 28,000-dalton proteins was observed in quiescent rat embryo fibroblasts following the addition of either the phorbol diester, phorbol-12-myristate-13 acetate, a calcium ionophore, A23187, or simply fresh serum. It is shown here that these two 28,000-dalton proteins are in fact two of the 28 kDa mammalian stress proteins.