Synthesis of docosahexaenoyl-, arachidonoyl- and palmitoyl-coenzyme A in ocular tissues.

The synthesis of long-chain acyl coenzyme A (CoA) was studied in the cornea, lens, vitreous, retina and pigment epithelium (PE) in the rat using [14C]-labeled palmitic, arachidonic and docosahexaenoic acids as substrates. Except for retina and PE, the ocular tissues studied showed relatively little enzyme activity with the fatty acid substrates. In addition, the enzyme activities were studied in homogenates and microsomal fractions from retina, pigment epithelial cells and choroid of frog, bovine and human eyes. Long-chain acyl CoA synthetase from the microsomal fraction exhibited three- to fivefold greater activity than homogenates in retina and PE. The enzyme activity was highest with palmitic acid, followed by arachidonic acid and docosahexaenoic acid. There were significant differences in enzyme activity between the species. The apparent Km (microM) and Vmax [nmol min-1 (mg protein)-1] values for the enzyme in bovine retinal microsomes were 7.91 +/- 0.39 (S.E.) and 21.6 +/- 1.04, respectively, for palmitic acid substrate and 5.88 +/- 0.25 and 4.58 +/- 0.21, respectively, for docosahexaenoic acid substrate. These values for bovine pigment epithelial microsomes were 13.0 +/- 0.27 and 36.9 +/- 1.18, respectively, for palmitic acid and 15.8 +/- 0.40 and 13.2 +/- 0.56, respectively, for docosahexaenoic acid. The synthesis of acyl CoA may play a central role in controlling the availability of free arachidonic acid for eicosanoid formation and in the retention of polyunsaturated fatty acid families (18:2, n-6 and 18:3, n-3) within cells of ocular tissues, particularly retina and retinal PE.