Differential effects of omega-3 fish oils on protein kinase activities in vitro.

We studied the in vitro effects of omega-3 fish oils and other fatty acids on the activity of crude protein kinase C from S49 lymphoma cells, on partially purified enzyme from rat cerebrum, on homogeneous protein kinase C from bovine brain, and, for comparison, on type I adenosine 3',5'-cyclic monophosphate (cAMP)-dependent protein kinase. In the absence of exogenous phospholipid, the fish oils cis-5,8,11,14,17-eicosapentaenoic acid (EPA) and acid (DCHA) enhance the catalytic cis-4,7,10,13,16,19-docosahexaenoic activity of protein kinase C and support the binding of [3H]phorbol 12,13-dibutyrate, both to approximately 50% of the level supported by phosphatidylserine. In the presence of phosphatidylserine, the omega-3 fatty acids reduce catalytic activity and [3H]phorbol 12,13-dibutyrate binding by about one-half.

Activity of an intracellular lymphocyte stimulator is independent of G-protein interactions, [Ca2+]i elevation, phosphoinositide hydrolysis, and protein kinase C translocation.

We have evaluated potential molecular mechanisms by which a group of synthetic lymphokine-like molecules, the 7,8-disubstituted guanine ribonucleosides, acts on second messenger pathways to augment the responses of murine B lymphocytes. Despite its extensive structural homology with GTP, 7-methyl-8-oxoguanosine (7-Me-8-oGuo), a prototypical disubstituted nucleoside, does not inhibit the binding of guanosine 5'-3-O-(thio)triphosphate either to purified G-proteins, or to G-proteins in situ in the plasma membrane. In contrast to anti-IgM antibodies, 7-Me-8-oGuo fails to induce elevation of intracellular free calcium in B lymphocytes.

Inhibition of phorbol ester binding and protein kinase C activity by heavy metals.

Other laboratories have reported biphasic effects of heavy metals on protein kinase C activity: stimulation followed by inhibition at higher concentrations. We demonstrate that these earlier findings most likely resulted from a combination of the effect of the heavy metals to liberate Ca2+ from Ca2+-EGTA buffer systems and the direct inhibitory effects of the metals on protein kinase C. Simulations of such interactions substantiate this conclusion.

Differences between human and goose erythrocytes in response to phorbol esters and expression of phorbol ester receptors.

Phorbol esters inhibited the uptake of a fluorescent glucose analogue in goose but not in human erythrocytes. Specific phorbol-12,13-dibutyrate (PDB) binding sites were identified in both goose and human erythrocytes. In the absence of Ca2+ and phospholipid, PDB binding in whole cell lysates was similar to that in intact cells, but addition of Ca2+ (0.

Asymmetric transport of a fluorescent glucose analogue by human erythrocytes.

A fluorescent glucose analogue, 6-deoxy-N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-aminoglucose (NBDG), was synthesized and its interactions with the hexose transport system of the human red blood cell were investigated. NBDG entry is inhibited by increasing concentrations of D-glucose (Ki = 2 mM). However, NBDG exit is unaffected by D-glucose in red blood cells.