Three major polypeptides of hepatitis B surface antigen (HBsAg), with mol. wt. 22000 (p22), 27000 (p27) and 68000 (p68), were separated by preparative SDS-PAGE. These three peptides as well as intact HBsAg were found to have almost identical amino acid compositions and carbohydrate was detected in p27 and p68 by PAS staining. Papain treatment of p68 produced two distinct peptides, p27 and p22. Moreover, when an artificial mixture of p27 and p22 in a ratio of 1:1 was treated with 0.2 m-periodate for 30 min at 37 °C, only p22 was detectable. These results suggest that p68 is composed of p27 and p22, and that p27 is a glycosylated product of p22. Thus, from the evidence obtained, it is possible that p22 (22000 peptide) is the minimum size of the unique hepatitis B virus (HBV) gene product involved.
CrestfieldA. M.,
MooreS.,
SteinW. H.1963; The preparation and enzymatic hydrolysis of reduced and S-carboxymethylated protein. Journal of Biological Chemistry 238:626–627
DejonghD. C.,
RadfordT.,
HribarJ. D.,
HanessianS.,
BieberM.,
DawsonG.,
SweeleyC. C.1969; Analysis of trimethylsilyl derivatives of carbohydrates by gas chromatography and mass spectrometry. Journal of the American Chemical Society 91:1728–1740
DreesmanG. R.,
GerinJ. L.1978; Structure of hepatitis virus. In Viral Hepatitis pp 645–648 Edited by
VyasG. N.,
CohenS. N.,
SchmidR.
Philadelphia: The Franklin Institute Press;
DreesmanG. R.,
ChairezR.,
SuarezM.,
HollingerF. B.,
CourtneyR. J.,
MelnickJ. L.1975; Production of antibody to individual polypeptides derived from purified hepatitis B surface antigen. Journal of Virology 16:508–515
LowryO. H.,
RosebroughN. J.,
FarrA. L.,
RandallR. J.1951; Protein measurement with the Folin phenol reagent. Journal of Biological Chemistry 193:265–275
PetersonD. L.,
RobertsI. M.,
VyasG. N.1977; Partial amino acid sequence of two major component polypeptides of hepatitis B surface antigen. Proceedings of the National Academy of Sciences of the United States of America 74:1530–1534
PeterssonG.1974; Gas chromatographic analysis of sugars and related hydroxy acids as acyclic oxime and ester trimethylsilyl derivatives. Carbohydrate Research 33:47–61
ShihJ. W.-K.,
GerinJ. L.1975; Immunochemistry of hepatitis B surface antigen (HBsAg): preparation and characterization of antibodies to the constituent polypeptides. Journal of Immunology 115:634639
ShihJ. W.-K.,
GerinJ. L.1977b; Proteins of hepatitis B surface antigen: amino acid compositions of the major polypeptides. Journal of Virology 21:1219–1222
ShiraishiH.,
ShirachiR.,
IshidaN.,
SekineT.1978; The specific reduction of a and d antigenic determinants after periodate treatment of hepatitis B surface antigen (HBsAg). Journal of General Virology 38:363–367
SkellyJ.,
HowardC. R.,
ZuckermanA. J.1978; The labelling of galactose residues in hepatitis B surface antigen glycoprotein. Journal of General Virology 41:447–457
SukenoN.,
ShiraishiH.,
IshidaN.1975; Conformational studies of hepatitis B surface antigen (HBsAg) by amino acid analysis. Tohoku Journal of Experimental Medicine 116:179–182
WeberK.,
OsbornM.1969; The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. Journal of Biological Chemistry 244:4406–4412