Hepatitis B surface antigen. Role of lipids in maintaining the structural and antigenic properties of protein components.
Most of the lipid components of hepatitis B surface antigen (HBsAg) can be removed by treatment with the non-ionic non-denaturing detergent beta-D-octyl glucoside (OG) followed by centrifugation through caesium chloride linear density gradients (density 1.15-1.32 g/ml). The conformational changes induced by the elimination of lipids decreased the helical content of HBsAg proteins from 52 to 28% as indicated by c.d. techniques. Measurements of the extent of quenching of protein fluorescence by iodide showed that half of the tryptophan residues which are buried in the native structure of HBsAg particles are brought close to the surface of the molecule by such conformational changes. The antigenic activity, as measured by binding to polyclonal antibodies, was decreased upon removal of lipids. Moreover, the six different antigenic sites recognized by our panel of monoclonal antibodies decreased their capacity to bind to the corresponding antibody when lipids were removed. However, the extent of this decrease differed for the different antibodies. Thus the apparent dependence of antibody binding on the lipid content seemed to indicate a greater involvement of the lipid-protein interaction for some of the epitopes than for others.
Gavilanes
F
F
Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias, Universidad Complutense, Madrid, Spain.
Gomez-Gutierrez
J
J
Aracil
M
M
Gonzalez-Ros
J M
JM
Ferragut
J A
JA
Guerrero
E
E
Peterson
D L
DL
D000911:Antibodies, Monoclonal / Q000276:immunology; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D006514:Hepatitis B Surface Antigens / Q000276:immunology* / Q000302:isolation & purification; D008055:Lipids / Q000032:analysis / Q000276:immunology* / Q000302:isolation & purification; D010758:Phosphorus / Q000032:analysis; D013050:Spectrometry, Fluorescence
D000911:Antibodies, Monoclonal; D006514:Hepatitis B Surface Antigens; D008055:Lipids; D010758:Phosphorus