Demonstration of phosphatidylinositol anchors on Ly-6 molecules by specific phospholipase C digestion and gel electrophoresis in octylglucoside.
Release by phosphatidylinositol-specific phospholipase C is frequently provided as evidence for membrane anchorage of a protein through phosphatidylinositol. Demonstration that the enzyme removes a lipophilic moiety from the protein is stronger evidence, and is presented here for members of the Ly-6 family of lymphocyte antigens: Ly-6A, C and E. Treatment of these molecules with the enzyme greatly increased their electrophoretic mobilities on polyacrylamide gels containing nonionic detergent. Furthermore, the mobilities of the digested, but not native Ly-6 molecules, were independent of detergent. This analytical method was applied to pure antigen, radiolabelled immunoprecipitate, or immunochemically detected Ly-6 antigens on blots.
Hammelburger
J W
JW
Memorial Sloan-Kettering Cancer Center, New York, N.Y. 10021.
Palfree
R G
RG
Sirlin
S
S
Hämmerling
U
U
D000818:Animals; D000950:Antigens, Ly / Q000032:analysis*; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D005960:Glucosides; D007124:Immunoenzyme Techniques; D051379:Mice; D008815:Mice, Inbred Strains; D010716:Phosphatidylinositols / Q000032:analysis*; D010738:Type C Phospholipases
D000950:Antigens, Ly; D003902:Detergents; D005960:Glucosides; D010716:Phosphatidylinositols; C018619:octyl-beta-D-glucoside; D010738:Type C Phospholipases