Adenylate kinase of Escherichia coli: evidence for a functional interaction in phospholipid synthesis.
Previous genetic and biochemical experiments have suggested that the adenylate kinase of Escherichia coli may be directly involved in phospholipid synthesis through formation of a complex with sn-glycerol-3-phosphate acyltransferase, the membrane-bound enzyme that catalyzes the first step in phospholipid synthesis. In this paper we report direct experiments to test this hypothesis. A mutation within the adenylate kinase structural gene is described that results in a temperature-sensitive phospholipid synthesis (assayed in vivo) and a temperature-sensitive acyltransferase. The adenylate kinase activity of this strain is only minimally altered either in vitro or [as assayed by adenosine 5'-triphosphate (ATP) levels] in vivo. This result demonstrates that the inhibition of phospholipid synthesis is not the result of reduced ATP levels. We report the purification of E. coli adenylate kinase to homogeneity; and find that the addition of homogeneous wild-type adenylate kinase to membranes containing a mutationally altered temperature-sensitive acyltransferase results in thermal stabilization of the acyltransferase activity. Ovalbumin has no such protective effect. Purified E. coli inner membranes contain several proteins that are precipitated by addition of anti adenylate kinase antibody to detergent-solubilized membranes.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000263:Adenylate Kinase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology; D004355:Drug Stability; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D005992:Glycerol-3-Phosphate O-Acyltransferase / Q000378:metabolism; D006358:Hot Temperature; D007158:Immunologic Techniques; D009154:Mutation; D010743:Phospholipids / Q000096:biosynthesis*; D010770:Phosphotransferases / Q000378:metabolism*
D010743:Phospholipids; D000255:Adenosine Triphosphate; D005992:Glycerol-3-Phosphate O-Acyltransferase; D010770:Phosphotransferases; D000263:Adenylate Kinase