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A transfer of (32P)-labeled phosphoryl groups has been demonstrated to occur between prelabeled Ehrlich cells (donor cells) and glioma cells (acceptor cells). The Ehrlich cells were generally prelabeled in vivo by an intraabdominal injection of (32P)-orthophosphate 24 hours prior to scarifice of the tumor-bearing mice. In some experiments two injections of (14C)-glucose were given to the animals in addition to the injection of (32P)-orthophosphate prior to sacrifice. The prelabeled Ehrlich cells were mixed with the glioma cells, that were still attached to the culture plates, and incubation took place in vitro. Labeled (32P)-phosphoryl (14C)-serine was isolated from the glioma cells as well as from the Ehrlich cells. The 32P/14C-ratio of (32P)-phosphoryl (14C)-serine of the glioma cells (acceptor cells) was significantly higher than found for the Ehrlich cells (donor cells). This was indicative of an interactive mechanism between the cells partly due to binding of membrane fragments from the Ehrlich cells on to the surface of the glioma cells and partly due to a phosphoryl group transfer between the two types of cells. Interaction experiments with a plasma membrane fraction prepared from the prelabeled Ehrlich cells vis à vis the glioma cells clearly showed that the phosphoryl group transfer mechanism between the cells was dominating and only a minor part of the interaction was due to adhesion. This transphosphorylation reaction represents a new type of interaction between cells. It was not influenced by either adenosine 3', 5'-monophosphate or adenosine 5'-diphosphate.

A phosphorylation of endogenous acceptor protein(s) has been demonstrated to occur in membraneous vesicles prepared from Ehrlich ascites tumour cells. The reaction was catalyzed by endogenous protein kinase in the presence of exogenous (gamma32P)ATP. A considerable increase of the specific protein kinase activity took place when the plasma membrane preparation was subjected to a further gradient centrifugation in Dextran T 150. This was done in the presence of a slightly alkaline phosphate buffer containing Mg-ions which resulted in the formation of a well defined vesicular preparation at density 1.035 in the gradient. The apparent Km and Vmax for the reaction with vesicles and exogenous (gamma32P)ATP were determined and found to be 0.022 mM and 0.23 nmol x mg-1 x 10 min-1, respectively. Neither cyclic AMP nor cyclic GMP did stimulate the protein kinase-catalyzed reaction. Instead, a clear inhibition of the reaction by the cyclic nucleotides was unexpectedly registered. Adenosine at 0.5 mM also inhibited the reaction. Calcium ions were inhibitory at all concentrations tested in the presence of a fixed (gamma32P)ATP/Mg2+ ratio. When Mg-ions were stoichiometrically replaced by Ca-ions practically no activity was observed.

A Mg2+- and Ca2+-dependent ATPase at the outer surface of human peripheral lymphocytes and hematopoietic cell lines has been studied. The enzyme activity varied about 60 fold between the extremes of the cell lines tested. There was no simple relationship between surface-ATPase activity and cell surface area. Nor was the presence of villi or any other morphological characteristic at the cell surface decisive for the degree of surface-ATPase activity. A correlation was, however, found between the ATPase activity and a phenomenon probably involving a contractile process namely immunoglobulin secretion, but it was not possible to establish an interdependence of these cell characteristics.