Ketone body and phosphoenolpyruvate formation by isolated hepatic mitochondria from Squalus acanthias (spiny dogfish)

Abstract

AbstractIn elasmobranchs, triglycerides are stored in the liver, which appears to be the primary site for lipolysis and oxidation of fatty acids, a major product being ketone bodies. The objective of this study was to obtain comparative information about properties of ketone body and phosphoenolpyruvate formation by liver of Squalus acanthias (spiny dogfish, a representative elasmobranch) by using an isolated hepatic mitochondrial system we have previously described for studying respiration and glutamine‐dependent citrulline synthesis. Acetate and palmitoyl‐CoA are stoichiometrically converted to ketone bodies at a rapid rate. The capability of mitochondria to convert short‐chain fatty acids to ketone bodies indicates that the previously reported inability of dogfish mitochondria to utilize short‐chain fatty acids to support respiration is not due to absence of nucleoside monophosphate kinase activity. At least two acyl‐CoA synthetase activities are present in mitochondrial extracts, one specific for acetate and another with specificity for longer‐chain fatty acids. Utilization of palmitoyl‐CoA is carnitine‐dependent and is inhibited by low concentrations of malonyl‐CoA. Pyruvate and alanine are also rapidly metabolized to ketone bodies. Phosphoenolpyruvate is formed at a significant rate from a number of substrates, including in particular aspartate and malate in the presence of α‐ketoglutarate. The osmolytes urea and trimethylamine oxide activate and inhibit, respectively, phosphoenolpyruvate formation; in contrast, these osmolytes inhibit and activate, respectively, ketone body and citrulline formation by isolated mitochondria. The results are consistent with the reported unique functions of dogfish liver in lipid metabolism and indicate that the basic enzymatic processes at the level of mitochondria appear to be analogous to those present in mammalian species.