Stability and Morphology Comparisons of Self-Assembled Virus-Like Particles from Wild-Type and Mutant Human Hepatitis B Virus Capsid Proteins

Margaret Newman, Fat Moon Suk, Maria Cajimat, Pong Kian Chua, Chiaho Shih

Research output: Contribution to journalArticle

54 Citations (Scopus)

Abstract

Instead of displaying the wild-type selective export of virions containing mature genomes, human hepatitis B virus (HBV) mutant I97L, changing from an isoleucine to a leucine at amino acid 97 of HBV core antigen (HBcAg), lost the high stringency of selectivity in genome maturity during virion export. To understand the structural basis of this so-called "immature secretion" phenomenon, we compared the stability and morphology of self-assembled capsid particles from the wild-type and mutant I97L HBV, in either full-length (HBcAg1-183) or truncated core protein contexts (HBcAg1-149 and HBcAg1-140). Using negative staining and electron microscopy, full-length particles appear as "thick-walled" spherical particles with little interior space, whereas truncated particles appear as "thin-walled" spherical particles with a much larger inner space. We found no significant differences in capsid stability between wild-type and mutant I97L particles under denaturing pH and temperature in either full-length or truncated core protein contexts. In general, HBV capsid particles (HBcAg1-183, HBcAg1-149, and HBcAg1-140) are very robust but will dissociate at pH 2 or 14, at temperatures higher than 75°C, or in 0.1% sodium dodecyl sulfate (SDS). An unexpected upshift banding pattern of the SDS-treated full-length particles during agarose gel electrophoresis is most likely caused by disulfide bonding of the last cysteine of HBcAg. HBV capsids are known to exist in natural infection as dimorphic T=3 or T=4 icosahedral particles. No difference in the ratio between T=3 (78%) and T=4 particles (20.3%) are found between wild-type HBV and mutant I97L in the context of HBcAg1-140. In addition, we found no difference in capsid stability between T=3 and T=4 particles successfully separated by using a novel agarose gel electrophoresis procedure.

Original languageEnglish
Pages (from-to)12950-12960
Number of pages11
JournalJournal of Virology
Volume77
Issue number24
DOIs
Publication statusPublished - Dec 2003
Externally publishedYes

Fingerprint

virus-like particles
Hepatitis B virus
Capsid Proteins
coat proteins
Virion
Capsid
mutants
capsid
Agar Gel Electrophoresis
Sodium Dodecyl Sulfate
Hepatitis B Core Antigens
Negative Staining
Temperature
sodium dodecyl sulfate
Isoleucine
virion
Human Genome
agarose
gel electrophoresis
Leucine

ASJC Scopus subject areas

  • Immunology

Cite this

Stability and Morphology Comparisons of Self-Assembled Virus-Like Particles from Wild-Type and Mutant Human Hepatitis B Virus Capsid Proteins. / Newman, Margaret; Suk, Fat Moon; Cajimat, Maria; Chua, Pong Kian; Shih, Chiaho.

In: Journal of Virology, Vol. 77, No. 24, 12.2003, p. 12950-12960.

Research output: Contribution to journalArticle

Newman, Margaret ; Suk, Fat Moon ; Cajimat, Maria ; Chua, Pong Kian ; Shih, Chiaho. / Stability and Morphology Comparisons of Self-Assembled Virus-Like Particles from Wild-Type and Mutant Human Hepatitis B Virus Capsid Proteins. In: Journal of Virology. 2003 ; Vol. 77, No. 24. pp. 12950-12960.
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