Reconstitution of hepatitis B surface antigen proteins into phospholipid vesicles.
Hepatitis B surface antigen (HBsAg), devoid of 75% of its total lipids has been reconstituted with several phospholipids by the detergent dialysis method, using the non-ionic detergent beta-D-octyl glucoside. Upon reconstitution with both neutral and acidic phospholipids, HBsAg particles had the same morphology and, as indicated by trypsin hydrolysis, the topology of the surface proteins was maintained. However, only negatively charged phospholipids were able to completely revert the conformational changes which had been induced by removal of the lipids. The helical content, as indicated by CD techniques, and the antigenic activity, as measured by binding to polyclonal antibodies, of HBsAg reconstituted with acidic phospholipids were practically identical to those of the native antigen. Cholesterol had no effect on the antigenic activity recovered by reconstitution with any of the phospholipids.
Gómez-Gutiérrez
J
J
Departamento de Bioquímica y Biología Molecular, Facultad de Ciencias Químicas, Universidad Complutense, Madrid, Spain.
Rodríguez-Crespo
I
I
Peterson
D L
DL
Gavilanes
F
F
D002308:Cardiolipins; D002942:Circular Dichroism; D003902:Detergents; D006514:Hepatitis B Surface Antigens / Q000737:chemistry* / Q000276:immunology / Q000648:ultrastructure; D006801:Humans; D006863:Hydrogen-Ion Concentration; D010713:Phosphatidylcholines; D010718:Phosphatidylserines; D010743:Phospholipids / Q000737:chemistry*; D011487:Protein Conformation
D002308:Cardiolipins; D003902:Detergents; D006514:Hepatitis B Surface Antigens; D010713:Phosphatidylcholines; D010718:Phosphatidylserines; D010743:Phospholipids