Glycoprotein Ib in the Triton-insoluble (cytoskeletal) fraction of blood platelets.
Glycoprotein Ib could be demonstrated in the Triton-insoluble (cytoskeletal) fraction of platelets prepared with EGTA by SDS-polyacrylamide gel electrophoresis and staining with the periodic acid Schiff's reagent. Crossed immunoelectrophoresis showed that glycoprotein Ib could be extracted from such Triton-insoluble residues when the extraction solution contained 1% Triton X-100 plus 5 mM CaCl2, but not if it also contained leupeptin. This indicates that glycoprotein Ib was associated to structures in the cytoskeletal fraction in such a way that it could be extracted only after activation of a calcium-dependent protease, and degradation of the actin-binding protein was demonstrated. After crossed immunoelectrophoresis of platelet extracts prepared in the presence of leupeptin or EDTA, a glycoprotein Ib-related, rocket-shaped immunoprecipitate was seen originating from the application well. This was interpreted as being related to glycoprotein Ib associated to actin polymers which did not sediment at low-speed centrifugation. Incubation of platelets with 32P as sodium phosphate led to incorporation of phosphatase-sensitive 32P in all of the glycoprotein Ib-related immunoprecipitates except for that of glycocalicin. This supports the idea that glycoprotein Ib traverses the plasma membrane and can be phosphorylated at the inner surface whereas glycocalicin represents the terminal part of the glycoprotein Ib alpha-chain exposed at the outer surface.
D001792:Blood Platelets / Q000032:analysis* / Q000648:ultrastructure; D003599:Cytoskeleton / Q000032:analysis; D004492:Edetic Acid; D006023:Glycoproteins / Q000097:blood*; D006801:Humans; D007123:Immunoelectrophoresis, Two-Dimensional; D007976:Leupeptins / Q000378:metabolism; D008565:Membrane Proteins / Q000097:blood*; D008970:Molecular Weight; D017830:Octoxynol; D010980:Platelet Membrane Glycoproteins; D011092:Polyethylene Glycols; D012995:Solubility; D013917:Thrombin / Q000494:pharmacology
D006023:Glycoproteins; D007976:Leupeptins; D008565:Membrane Proteins; D010980:Platelet Membrane Glycoproteins; D011092:Polyethylene Glycols; D017830:Octoxynol; D004492:Edetic Acid; D013917:Thrombin; C032854:leupeptin