cerulenin and 2-aminobicyclo(2-2-1)heptane-2-carboxylic-acid

To determine whether protein acylation plays a role in the effects of glucose on the insulin secreting β-cell.. The measurement of (3)H-palmitate incorporation into protein in the INS 832/13 cell that has a robust and well-characterized biphasic insulin secretory response to stimulation with glucose.. Stimulating the cells with glucose increased the incorporation of (3)H-palmitic acid into protein by up to 90%. Similarly, 2-aminobicyclo [2.2.1] heptane-2-carboxylic acid (BCH) the non-metabolizable analog of leucine that mimics the stimulatory effect of glucose on insulin secretion also increased the incorporation of (3)H-palmitic acid into protein. Treatment of cell lysates with hydroxylamine substantially reduced the incorporation indicating that most of the incorporation was due to enzymatic palmitoylation of proteins. Cerulenin, a classical inhibitor of protein acylation also substantially reduced the incorporation. Using PAGE and autoradiography a glucose-induced increase in protein palmitoylation and specific glucose-induced increases in the palmitoylation of proteins of 30, 44, 48 and 76kD were identified.. The data suggest that protein acylation plays multiple roles in β-cell function.

Topics: Acylation; Amino Acids, Cyclic; Animals; Autoradiography; Cells, Cultured; Cerulenin; Electrophoresis, Polyacrylamide Gel; Glucose; Hydroxylamine; Insulin-Secreting Cells; Palmitic Acid; Proteins; Rats