Recombinant hepatitis B surface antigen and anionic phospholipids share a binding region in the fifth domain of beta2-glycoprotein I (apolipoprotein H).
Human beta2-glycoprotein I (beta 2GPI) binds to recombinant hepatitis B surface antigen (rHBsAg), but the location of the binding domain on beta 2GPI is unknown. It has been suggested that the lipid rather than the protein moiety of rHBsAg binds to beta 2GPI. Since beta 2GPI binds to anionic phospholipids (PL) through its lipid-binding region in the fifth domain of beta 2GPI, we predicted that this lipid-binding region may also be involved in binding rHBsAg. In this study, we examined rHBsAg binding to two naturally occurring mutants of beta 2GPI, Cys306Gly and Trp316Ser, or evolutionarily conserved hydrophobic amino acid sequence, Leu313-Ala314-Phe315 in the fifth domain of beta 2GPI. The two naturally occurring mutations and two mutagenized amino acids, Leu313Gly or Phe315Ser, disrupted the binding of recombinant beta 2GPI (rbeta 2GPI) to both rHBsAg and cardiolipin (CL), an anionic PL. These results suggest that rHBsAg and CL share the same region in the fifth domain of beta2GPI. Credence to this conclusion was further provided by competitive ELISA, where CL-bound rbeta 2GPI was incubated with increasing amounts of rHBsAg. As expected, pre-incubation of rbeta 2GPI with CL precluded binding to rHBsAg, indicating that CL and rHBsAg bind to the same region on beta 2GPI. Our data provide evidence that the lipid (PL) rather than the protein moiety of rHBsAg binds to beta 2GPI and that this binding region is located in the fifth domain of beta 2GPI, which also binds to anionic PL.
Mehdi
Haider
H
Department of Human Genetics, Graduate School of Public Health, University of Pittsburgh, Pittsburgh, PA 15261, USA. mehdih@upmc.edu
Naqvi
Asma
A
Kamboh
M Ilyas
MI
D000595:Amino Acid Sequence; D026922:Amino Acid Transport Systems, Acidic / Q000378:metabolism; D001665:Binding Sites; D001667:Binding, Competitive; D002308:Cardiolipins / Q000378:metabolism; D006514:Hepatitis B Surface Antigens / Q000378:metabolism*; D066298:In Vitro Techniques; D016297:Mutagenesis, Site-Directed; D010743:Phospholipids / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000378:metabolism; D053482:beta 2-Glycoprotein I / Q000235:genetics / Q000378:metabolism*
D026922:Amino Acid Transport Systems, Acidic; D002308:Cardiolipins; D006514:Hepatitis B Surface Antigens; D010743:Phospholipids; D011994:Recombinant Proteins; D053482:beta 2-Glycoprotein I