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J. Biol. Chem., Vol. 280, Issue 23, 21713-21719, June 10, 2005

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Hepatitis B Virus DNA-negative Dane Particles Lack Core Protein but Contain a 22-kDa Precore Protein without C-terminal Arginine-rich Domain^*

Tatsuji Kimura, Nobuhiko Ohno¶, Nobuo Terada¶, Akinori Rokuhara||, Akihiro Matsumoto||, Shintaro Yagi, Eiji Tanaka||, Kendo Kiyosawa||, Shinichi Ohno¶, and Noboru Maki

From the Research and Development Division, Advanced Life Science Institute, Inc., Wako, Saitama 351-0112, Japan, the ^¶Department of Anatomy, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, Tamaho-cho, Yamanashi 409-3898, Japan, and the ^||Department of Internal Medicine, Shinshu University School of Medicine, Matsumoto, Nagano 390-8621, Japan

DNA-negative Dane particles have been observed in hepatitis B virus (HBV)-infected sera. The capsids of the empty particles are thought to be composed of core protein but have not been studied in detail. In the present study, the protein composition of the particles was examined using new enzyme immunoassays for the HBV core antigen (HBcAg) and for the HBV precore/core proteins (core-related antigens, HBcrAg). HBcrAg were abundant in fractions slightly less dense than HBcAg and HBV DNA. Three times more Dane-like particles were observed in the HBcrAg-rich fraction than in the HBV DNA-rich fraction by electron microscopy. Western blots and mass spectrometry identified the HBcrAg as a 22-kDa precore protein (p22cr) containing the uncleaved signal peptide and lacking the arginine-rich domain that is involved in binding the RNA pregenome or the DNA genome. In sera from 30 HBV-infected patients, HBcAg represented only a median 10.5% of the precore/core proteins in enveloped particles. These data suggest that most of the Dane particles lack viral DNA and core capsid but contain p22cr. This study provides a model for the formation of the DNA-negative Dane particles. The precore proteins, which lack the arginine-rich nucleotide-binding domain, form viral RNA/DNA-negative capsid-like particles and are enveloped and released as empty particles.

Received for publication, February 10, 2005 , and in revised form, March 31, 2005.

^* The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: Research and Development Division, Advanced Life Science Institute, Inc., 2-10-23 Maruyamadai, Wako, Saitama 351-0112, Japan. Tel.: 81-48-465-2761; Fax: 81-48-465-2765; E-mail: tkimura{at}alsi-i.co.jp.

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				D. K.-H. Wong, Y. Tanaka, C.-L. Lai, M. Mizokami, J. Fung, and M.-F. Yuen Hepatitis B Virus Core-Related Antigens as Markers for Monitoring Chronic Hepatitis B Infection J. Clin. Microbiol., December 1, 2007; 45(12): 3942 - 3947. [Abstract] [Full Text] [PDF]