Evidence for structural differences in the S domain of L in comparison with S protein of hepatitis B virus.
The structures of the large (L), middle (M) and small (S) versions of the envelope proteins of hepatitis B virus remain poorly characterized due to the complex nature of their conformations. Several groups have proposed transmembrane topological models depicting the lumenally and cytosolically disposed regions of these proteins. Recently, post-translational topological changes in L have been described. However, no overall differences in the topology of the S domains of the L or M, to the S protein are predicted. In this report, we investigated a previously uncharacterized anti-S monoclonal antibody (MAb), 6B1, which recognizes a conformation-sensitive epitope in S. Unlike other anti-S MAbs tested, this MAb did not recognize its epitope in the S domain of L protein. Interestingly, however, the M protein was efficiently recognized. This unique characteristic of MAb 6B1 has allowed us to study the intracellular distribution of L and S proteins. In cells expressing both L and S, L re-localized from the endoplasmic reticulum-Golgi intermediate compartment (ERGIC) to the membrane-associated distribution of S protein indicating that L and S interact with each other. This was confirmed by immunoprecipitation assays, which also showed that the interaction between L and S results in the secretion of L protein from cells. Overall, the ability of MAb 6B1 to selectively recognize S and M, but not L, strongly points to the existence of significant topological differences in the S domain of L. The availability of this important reagent should help further our understanding of the structure of HBV surface antigens.
Clayton
Reginald F
RF
MRC Virology Unit, Institute of Virology, Church Street, Glasgow G11 5JR, UK1.
Owsianka
Ania
A
MRC Virology Unit, Institute of Virology, Church Street, Glasgow G11 5JR, UK1.
Patel
Arvind H
AH
MRC Virology Unit, Institute of Virology, Church Street, Glasgow G11 5JR, UK1.
D000911:Antibodies, Monoclonal / Q000276:immunology; D002460:Cell Line; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005822:Genetic Vectors; D006056:Golgi Apparatus / Q000378:metabolism; D006514:Hepatitis B Surface Antigens / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D006515:Hepatitis B virus / Q000276:immunology*; D018613:Microscopy, Confocal; D011233:Precipitin Tests; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000276:immunology; D014616:Vaccinia virus / Q000235:genetics; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D006514:Hepatitis B Surface Antigens; C096362:L protein, hepatitis B virus; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins