Cholinesterase solubilizing factor from Cytophaga sp. is a phosphatidylinositol-specific phospholipase C.
In the culture supernatant of Cytophaga sp. we detected an enzyme that converted glycosylphosphatidyl-inositol-anchored acetylcholinesterase to the hydrophilic form. This enzyme had a cleavage specificity of a phospholipase C. It hydrolyzed phosphatidylinositol but did not act on phosphatidylcholine. On gel filtration the enzyme migrated with an apparent molecular mass of about 17 kDa. It displayed maximal activity between pH 6-6.5 and did not require cofactors for the expression of catalytic activity. Mercurials and zinc ions inhibited the enzyme and its activity also decreased with increasing ionic strength in the assay. With acetylcholinesterase as substrate optimal activity was obtained in pure micelles of Triton X-100, whereas in mixed micelles containing Triton X-100 and phosphatidylcholine the activity was reduced. The enzyme from Cytophaga sp. showed little activity towards acetylcholinesterase embedded in intact membranes where more than 1000-times higher concentrations of phosphatidylinositol-specific phospholipase C was necessary to solubilize acetylcholinesterase as compared to acetylcholinesterase in detergent micelles.
Jäger
K
K
Institut für Biochemie und Molekularbiologie, Universität Bern, Switzerland.
Stieger
S
S
Brodbeck
U
U
D000110:Acetylcholinesterase / Q000378:metabolism*; D015153:Blotting, Western; D002412:Cations; D002802:Cholinesterases / Q000378:metabolism*; D003429:Cross Reactions; D003589:Cytophaga / Q000201:enzymology*; D003902:Detergents; D006863:Hydrogen-Ion Concentration; D008823:Micelles; D008970:Molecular Weight; D017830:Octoxynol; D010716:Phosphatidylinositols / Q000378:metabolism*; D011092:Polyethylene Glycols; D012995:Solubility; D013379:Substrate Specificity; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism*
D002412:Cations; D003902:Detergents; D008823:Micelles; D010716:Phosphatidylinositols; D011092:Polyethylene Glycols; D017830:Octoxynol; D000110:Acetylcholinesterase; D002802:Cholinesterases; D010738:Type C Phospholipases