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Metabolism and Enzymology

Free Fatty Acids Regulate Two Galactosyltransferases in Chloroplast Envelope Membranes Isolated from Spinach Leaves

Division of Environmental Biology, The National Institute for Environmental Studies, Onogawa, Tsukuba, Ibaraki 305, Japan, Department of Biology, University of Tokyo, Komaba, Meguro-ku, Tokyo 153, Japan

Effects of MgCl₂ and free fatty acids (FFA) on galactolipid:galactolipid galactosyltransferase (GGGT) and UDP-galactose: 1,2-diacylglycerol galactosyltransferase (UDGT) in chloroplast envelope membranes isolated from spinach (Spinacia oleracea L.) leaves were examined. GGGT activity was sigmoidally stimulated by MgCl₂ with a saturated concentration of more than 5 millimolar. Free -linolenic acid (18:3) caused a drastic increase in GGGT activity under limiting concentrations of MgCl₂, without affecting its maximum activity at higher MgCl₂ concentrations. Free 18:3 alone did not affect the GGGT activity. The effective species of FFA for the stimulation of GGGT activity in the presence of MgCl₂ were unsaturated 16- and 18-carbon fatty acids. GGGT activity was also stimulated by 18:3 in the presence of MnCl₂, CaCl₂ and a high concentration of KCl in place of MgCl₂. UDGT activity was hyperbolically enhanced by MgCl₂ with a saturated concentration of 1 to 2 millimolar. In contrast to GGGT, UDGT was severely inhibited by 18:3, and MgCl₂-induced stimulation was completely abolished by 18:3. Unsaturated 16- and 18-carbon fatty acids were more inhibitory to UDGT than the saturated acids. The dependence of GGGT activity on monogalactosyldiacylglycerol (MGDG) and MgCl₂ concentrations was identical in the envelope membranes isolated from non- and ozone (0.5 microliter/liter)-fumigated spinach leaves, indicating that GGGT remained active in the leaves during ozone fumigation. The results are discussed in relation to the regulation of galactolipid biosynthesis by the endogenous FFA in the envelopes and to the involvement of GGGT in the triacylglycerol synthesis from MGDG in ozone-fumigated leaves.

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