Purification and structural analysis of the hepatitis B virus preS1 expressed from Escherichia coli.
The preS1 of hepatitis B virus (HBV) is located at the outermost part of the envelope protein and possesses several functionally important regions such as hepatocyte receptor-binding site and virus-neutralizing epitopes. As the first step to understand the structure-function relationship for the preS1 antigen, we have purified the preS1 and performed its structural characterization by circular dichroism (CD) spectroscopy. The preS1 was purified to near homogeneity from bacterially expressed glutathione S-transferase (GST)-preS1 fusion protein by two-step purification, affinity chromatography on glutathione-agarose column, and cation-exchange chromatography on Mono S column. The CD analysis showed that the purified preS1, which was largely unstructured in aqueous solution, acquired a significant (16%) alpha-helical structure when analyzed in 50% trifluoroethanol or 20 mM SDS. The results suggest that the preS1 assumes a mainly unstructured conformation and may form induced secondary structures upon binding to target proteins or under hydrophobic environment.
Maeng
C Y
CY
Antibody Engineering Research Laboratory, Korea Research Institute of Bioscience and Biotechnology, Yusong, Taejon, 305-600.
Oh
M S
MS
Park
I H
IH
Hong
H J
HJ
D000595:Amino Acid Sequence; D001483:Base Sequence; D002942:Circular Dichroism; D004926:Escherichia coli / Q000235:genetics; D006514:Hepatitis B Surface Antigens / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D006515:Hepatitis B virus / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010957:Plasmids / Q000235:genetics; D011498:Protein Precursors / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification
D006514:Hepatitis B Surface Antigens; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; C050806:presurface protein 1, hepatitis B surface antigen