Binding of cardiolipin to polystyrene beads: evidence for a lamellar phase orientation.
The association of cardiolipin with polystyrene beads was studied using 31P-NMR and electron microscopy. In the presence and absence of fetal calf serum, cardiolipin appeared to bind to the polystyrene beads in lamellar phase as assessed by 31P-NMR imaging. Electron microscopic analysis revealed an even coating of phospholipid about the beads with extensive micelle binding. Cardiolipin-coated beads challenged with ACA-positive sera followed by immunogold indicated antibody bound to micelles associated with the bead. Studies conducted with ACA IgG purified from patient sera indicated that some ACA bound to CL beads in the absence of a source of ACA cofactor (i.e. gelatin-blocked beads), some ACA required beta 2-GPI for binding (i.e. no binding in the presence of beta 2-GPI-depleted plasma), whereas other ACA which showed negligible binding with gelatin-blocked beads, showed enhanced binding in the presence of beta 2-GPI-depleted plasma. The data indicate that: (1) cardiolipin binds to polystyrene beads in lamellar phase, (2) ACA bind to phospholipid micelles bound directly to the polystyrene beads, and (3) ACA differ between individuals displaying varying phospholipid and phospholipid/cofactor substrate specificities.
Stewart
M W
MW
Department of Laboratory Medicine and Pathology, University of Alberta Hospitals, Edmonton, Canada.
Gordon
P A
PA
Etches
W S
WS
Marusyk
H
H
Poppema
S
S
Bigam
C
C
Sykes
B
B
D017153:Antibodies, Anticardiolipin / Q000378:metabolism; D002308:Cardiolipins / Q000378:metabolism*; D005434:Flow Cytometry; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D011137:Polystyrenes / Q000378:metabolism*
D017153:Antibodies, Anticardiolipin; D002308:Cardiolipins; D006023:Glycoproteins; D011137:Polystyrenes