title: Antibodies prepared to Bacillus cereus phospholipase C crossreact with a phosphatidylcholine preferring phospholipase C in mammalian cells.
abstract: Antibodies against Bacillus cereus phospholipase C were prepared in rabbits and used to affinity purify a phosphatidylcholine-preferring phospholipase C from a human monocytic cell line. Affinity chromatography resulted in an approximately 3000-fold, one-step enrichment of phospholipase C. The human enzyme had an apparent molecular mass of 40,000 daltons as determined by SDS gel electrophoresis. Western blotting analysis demonstrated that this protein interacted specifically with the rabbit antibody raised against bacterial phospholipase C. The purified enzyme preferred phosphatidylcholine as a substrate, was neutral pH active and was inhibited by EGTA. These studies demonstrate that antibodies raised against bacterial phospholipase C may be useful in purifying phospholipase C from a human source.
mesh_terms: D000906:Antibodies / Q000276:immunology; D001409:Bacillus cereus / Q000201:enzymology; D002846:Chromatography, Affinity; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D010713:Phosphatidylcholines / Q000378:metabolism*; D013379:Substrate Specificity; D010738:Type C Phospholipases / Q000276:immunology / Q000302:isolation & purification*
chemical_list: D000906:Antibodies; D010713:Phosphatidylcholines; D010738:Type C Phospholipases