Degraded and stable phosphatidylglycerol in Escherichia coli inner and outer membranes, and recycling of fatty acyl residues.

ABSTRACT

The metabolic fate of membrane phospholipids in exponentially growing Escherichia coli was reexamined by incorporation and chase of labeled precursors [32P]phosphate, [2-3H]glycerol and 3H-labeled fatty acids. It was found that the well-known turnover of phosphatidylglycerol lasted only about two generation times; after which period, the remaining labeled phosphatidylglycerol, approximately one-third of the total, was stable for at least the subsequent two generation times. The location of the stable phosphatidylglycerol pool remaining after the turnover in the outer and inner membrane was investigated. Both envelopes were found to contain stable phosphatidylglycerol so that the existence of a stable portion cannot be ascribed to its exclusive location in one leaflet. In some experiments, a small loss of labeled phosphatidylethanolamine was also observed, and upon fractionation this was found to occur exclusively in the outer membrane. [32P]Phosphate and [2-3H]glycerol labels of the degraded phospholipids were lost from lipid-soluble material, whereas labeled fatty acid, palmitate or oleate was reincorporated into newly synthesized phosphatidylethanolamine and phosphatidylglycerol, so that total fatty acid label remained constant in (membrane) phospholipid during chase. In view of the amount of glycerol lost, the recycling of the fatty acids under the form of diacylglycerols to phosphatidic acid does not appear to be the predominant pathway of reincorporation. After double labeling with [32P]phosphate and [3H]palmitate, followed by chase, a complete balance sheet of loss and reincorporation of fatty acid, in the three phospholipids, in the two envelopes could be established. Results indicate that fatty acid was reincorporated essentially in the inner membrane phospholipids. Movements of phospholipids and of fatty acids from one membrane to another and in the plane of each layer are discussed in the light of the results.