Purification and characterization of an inhibitor of phospholipase A1 in Bacillus subtilis.
The protoplasts of a mutant of Bacillus subtilis 168 (B. subtilis CMK33) are osmotically fragile when compared to protoplasts of the parent organism and contain an active, membrane-associated phospholipase A1. A protein found in the parent organism specifically inhibits the phospholipase A1 (Kent, C., and Lennarz, W.J. (1972) Proc. Nat. Acad. Sci. U.S.A. 69, 2793-2797). The inhibitor exists in both a soluble and particulate form. The soluble inhibitor is not found in the cytoplasm, but rather in a "periplasmic" fraction released from the cell during incubation with lysozyme. The soluble inhibitor has been purified to homogeneity by diethylaminoethyl-cellulose and hydroxylapatite chromatography. Its molecular weight is 28,000 to 32,000 as determined by gel filtration chromatography and 36,000 to 37,000 as determined by sodium dodecyl sulfate-urea gel electrophoresis. The inhibitor appears to inactivate the membrane bound phospholipase A1 by an enzymatic process that is dependent on time and protein concentration. Binding of the inhbitor  to the membrane-associated phospholipase cannot be detected. When purified inhibitor is added to cells of B. subtilis CMK33 during treatment with lysozyme, the osmotic stability of the resultant protoplasts is similar to that of protoplasts of the wild type of organism.
D001412:Bacillus subtilis / Q000032:analysis / Q000201:enzymology* / Q000502:physiology; D001426:Bacterial Proteins / Q000302:isolation & purification / Q000502:physiology*; D001665:Binding Sites; D002462:Cell Membrane / Q000032:analysis; D002845:Chromatography; D004492:Edetic Acid / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D008970:Molecular Weight; D009154:Mutation; D009994:Osmolar Concentration; D010740:Phospholipases / Q000037:antagonists & inhibitors*; D011485:Protein Binding; D013347:Subcellular Fractions / Q000032:analysis; D013395:Sucrose / Q000494:pharmacology
D001426:Bacterial Proteins; D013395:Sucrose; D004492:Edetic Acid; D010740:Phospholipases