The biosynthetic incorporation of diacetylenic fatty acids into the biomembranes of Acholeplasma laidlawii A cells and polymerisation of the biomembranes by irradiation with ultraviolet light.

Acholeplasma laidlawii A has been grown in media containing synthetic, long chain C20- and C23-fatty acids possessing a diacetylene group in their acyl chains. Growth on the C23 diacetylenic acid was poor but was good on the C20 acid. Biosynthetic incorporation of the fatty acids occurs; as much as 90% of the membrane lipid fatty acyl chains consisting of the C20-diacetylenic fatty acid, the remainder being shorter chain, saturated fatty acids. The thermal phase transition of this biomembrane has been studied and a differential scanning calorimetry heating curve shows the presence of an endotherm corresponding to a membrane lipid phase transition occurring at about 26 degrees C. The lipid class composition of membranes containing the C20-diacetylene lipids was examined and found to be similar to membranes from cells grown on oleic acid-containing medium. (The ratio of monoglucosyl- to diglucosyldiacylglycerols was the same but the ratio of glycolipid to phosphatidylglycerol was higher in the cells grown with diacetylene fatty acids). Upon irradiation with ultraviolet light the cells and isolated biomembranes become coloured, either red or yellow depending upon their thermal history. The colour change indicates that extensive cross-linking of the lipids of the biomembranes of A. laidlawii has occurred and that a conjugated polymeric structure has been formed. Analysis of the extracted lipids from the biomembranes by GLC indicates that extensive cross-linking of the lipid chains within the biomembrane of a natural cell system has been achieved. The monoglucosyldiacylglycerols cross-link more readily that do the phosphatidylglycerol lipids. The effect of such lipid cross-linking or polymerisation on the activity at 35 degrees C of an intrinsic membrane-bound enzyme, NADH oxidase, and ribonuclease, an extrinsic membrane-bound enzyme, was studied. The NADH oxidase activity decreased rapidly upon cross-linking of the lipid environment whereas ribonuclease activity was unaffected. The potential for future studies of polymerised model and natural biomembranes is discussed.