Structure of the Hepatitis A Virion: Peptide Mapping of the Capsid Region

Overview

Journal J Virol

Publisher American Society for Microbiology

Date 1986 May 1

PMID 3009847

Citations 9

Authors

C M Wheeler

B H Robertson

G Van Nest

D Dina

D W Bradley

H A Fields

Affiliations

Soon will be listed here.

Abstract

Milligram amounts of highly purified hepatitis A virus (HAV) were obtained from persistently infected cell cultures. The HAV polypeptides were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to nitrocellulose for detection by an enzyme-linked immunotransfer blot procedure. The HAV nucleotide-derived amino acid sequence was subjected to computer analysis to identify potential immunogenic regions within the HAV capsid polypeptides. Synthetic peptides corresponding to selected regions of each of the larger putative capsid polypeptides were coupled to keyhole limpet hemocyanin and used to immunize rabbits. Four of six anti-HAV peptide sera were strongly reactive. Antipeptide serum generated against amino acids (a.a.) 75 through 82 reacted with the 27,000-molecular-weight (MW) polypeptide; serum against a.a. 279 through 285 reacted with the 29,000-MW HAV polypeptide; and sera against a.a. 591 through 602 and 606 through 618 reacted with the 33,000-MW HAV polypeptide. These reactions enabled the identification of the gene order of the larger HAV P1 region gene products. Our data indicate the following molecular weights: HAV VP2 or 1B, 27,000; HAV VP3 or 1C, 29,000; and HAV VP1 or 1D, 33,000.

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References

Coulepis A, Tannock G, Locarnini S, Gust I . Evidence that the genome of hepatitis A virus consists of single-stranded RNA. J Virol. 1981; 37(1):473-7. PMC: 171023. DOI: 10.1128/JVI.37.1.473-477.1981. View

GREENWOOD F, Hunter W, GLOVER J . THE PREPARATION OF I-131-LABELLED HUMAN GROWTH HORMONE OF HIGH SPECIFIC RADIOACTIVITY. Biochem J. 1963; 89:114-23. PMC: 1202279. DOI: 10.1042/bj0890114. View

Adams J . Heavy metal intensification of DAB-based HRP reaction product. J Histochem Cytochem. 1981; 29(6):775. DOI: 10.1177/29.6.7252134. View

von der Helm K, Winnacker E, Deinhardt F, Frosner G, Gauss-Muller V, Bayerl B . Cloning of hepatitis A virus genome. J Virol Methods. 1981; 3(1):37-43. DOI: 10.1016/0166-0934(81)90020-3. View

Hopp T, Woods K . Prediction of protein antigenic determinants from amino acid sequences. Proc Natl Acad Sci U S A. 1981; 78(6):3824-8. PMC: 319665. DOI: 10.1073/pnas.78.6.3824. View

Rueckert R, Pallansch M . Preparation and characterization of encephalomyocarditis (EMC) virus. Methods Enzymol. 1981; 78(Pt A):315-25. View

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

Marglin A, MERRIFIELD R . Chemical synthesis of peptides and proteins. Annu Rev Biochem. 1970; 39:841-66. DOI: 10.1146/annurev.bi.39.070170.004205. View

Feinstone S, Kapikian A, Purceli R . Hepatitis A: detection by immune electron microscopy of a viruslike antigen associated with acute illness. Science. 1973; 182(4116):1026-8. DOI: 10.1126/science.182.4116.1026. View

10.

Swaney J, Woude G, Bachrach H . Sodium dodecylsulfate-dependent anomalies in gel electrophoresis: alterations in the banding patterns of foot-and-mouth disease virus polypeptides. Anal Biochem. 1974; 58(2):337-46. DOI: 10.1016/0003-2697(74)90201-2. View

11.

Liu F, Zinnecker M, Hamaoka T, Katz D . New procedures for preparation and isolation of conjugates of proteins and a synthetic copolymer of D-amino acids and immunochemical characterization of such conjugates. Biochemistry. 1979; 18(4):690-3. DOI: 10.1021/bi00571a022. View

12.

Guesdon J, Ternynck T, Avrameas S . The use of avidin-biotin interaction in immunoenzymatic techniques. J Histochem Cytochem. 1979; 27(8):1131-9. DOI: 10.1177/27.8.90074. View

13.

Kew O, Pallansch M, Omilianowski D, Rueckert R . Changes in three of the four coat proteins of oral polio vaccine strain derived from type 1 poliovirus. J Virol. 1980; 33(1):256-63. PMC: 288542. DOI: 10.1128/JVI.33.1.256-263.1980. View

14.

Green N, Alexander H, Olson A, Alexander S, Shinnick T, Sutcliffe J . Immunogenic structure of the influenza virus hemagglutinin. Cell. 1982; 28(3):477-87. DOI: 10.1016/0092-8674(82)90202-1. View

15.

Tsang V, Peralta J, Simons A . Enzyme-linked immunoelectrotransfer blot techniques (EITB) for studying the specificities of antigens and antibodies separated by gel electrophoresis. Methods Enzymol. 1983; 92:377-91. DOI: 10.1016/0076-6879(83)92032-3. View

16.

Gust I, Coulepis A, Feinstone S, Locarnini S, Moritsugu Y, Najera R . Taxonomic classification of hepatitis A virus. Intervirology. 1983; 20(1):1-7. DOI: 10.1159/000149367. View

17.

Ticehurst J, Racaniello V, Baroudy B, Baltimore D, Purcell R, Feinstone S . Molecular cloning and characterization of hepatitis A virus cDNA. Proc Natl Acad Sci U S A. 1983; 80(19):5885-9. PMC: 390180. DOI: 10.1073/pnas.80.19.5885. View

18.

Tsang V, Hancock K, MADDISON S, Beatty A, Moss D . Demonstration of species-specific and cross-reactive components of the adult microsomal antigens from Schistosoma mansoni and S. japonicum (MAMA and JAMA). J Immunol. 1984; 132(5):2607-13. View

19.

Rueckert R, Wimmer E . Systematic nomenclature of picornavirus proteins. J Virol. 1984; 50(3):957-9. PMC: 255765. DOI: 10.1128/JVI.50.3.957-959.1984. View

20.

Bradley D, Schable C, McCaustland K, Cook E, Murphy B, Fields H . Hepatitis A virus: growth characteristics of in vivo and in vitro propagated wild and attenuated virus strains. J Med Virol. 1984; 14(4):373-86. DOI: 10.1002/jmv.1890140410. View