Characterization of a slow-migrating component of the rabies virus matrix protein strongly associated with the viral glycoprotein

被引:3
作者
Nakahara, T [1 ]
Toriumi, H [1 ]
Irie, T [1 ]
Takahashi, T [1 ]
Ameyama, S [1 ]
Mizukoshi, M [1 ]
Kawai, A [1 ]
机构
[1] Kyoto Univ, Grad Sch Pharmaceut Sci, Dept Mol Microbiol, Sakyo Ku, Kyoto 6068501, Japan
关键词
rabies virus; matrix protein; conformational change; monoclonal antibody #3-9-16; dimer formation;
D O I
10.1111/j.1348-0421.2003.tb03458.x
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
We investigated multiple forms of rabies virus matrix (M) protein. Under non-reducing electrophoretic conditions, we detected, in addition to major bands of monomer forms (23- and 24-kDa) of M protein, an M antigen-positive slow-migrating minor band (about 54 kDa) in both the virion and infected cells. Relative contents of the 54-kDa and monomer components in the virion were about 20-30% and 70-80% of the whole M protein, respectively, while the content of the 54-kDa component was smaller (about 10-20% of the total M protein) in the cell than in the virion. The 54-kDa components could be extracted from the infected cells with sodium deoxycholate, but they were quite resistant to extraction with 1% nonionic detergents by which most monomer components were solubilized. The 54-kDa component was precipitated more efficiently than the monomer by a monoclonal antibody (mAb; #3-9-16), which recognized a linear epitope located at the N-terminal of the M protein. The mAb #3-9-16 coprecipitated the viral glycoprotein (G), which was demonstrated to be due to strong association between the G and 54-kDa component of the M protein. Monomers and the 54-kDa polypeptide migrated to the same isoelectric point (pI) in two-dimensional (2-D) gel electrophoresis, implicating that the 54-kDa component was composed of component(s) of the same pI as that of the M protein monomers. From these results, we conclude that the M antigen-positive 54-kDa polypeptide is a homodimer of M protein, taking an N-terminal-exposed conformation, and is strongly associated with the viral glycoprotein. Possible association with a membrane microdomain of the cell will be discussed.
引用
收藏
页码:977 / 988
页数:12
相关论文
共 41 条
[1]   Monoclonal antibody #3-9-16 recognizes one of the two isoforms of rabies virus matrix protein that exposes its N-terminus on the virion surface [J].
Ameyama, S ;
Toriumi, H ;
Takahashi, T ;
Shimura, Y ;
Nakahara, T ;
Honda, Y ;
Mifune, K ;
Uchiyama, T ;
Kawai, A .
MICROBIOLOGY AND IMMUNOLOGY, 2003, 47 (09) :639-651
[2]   Lipid raft microdomains: A gateway for compartmentalized trafficking of Ebola and Marburg viruses [J].
Bavari, S ;
Bosio, CM ;
Wiegand, E ;
Ruthel, G ;
Will, AB ;
Geisbert, TW ;
Hevey, M ;
Schmaljohn, C ;
Schmaljohn, A ;
Aman, MJ .
JOURNAL OF EXPERIMENTAL MEDICINE, 2002, 195 (05) :593-602
[3]   ROLE OF MATRIX PROTEIN IN CYTOPATHOGENESIS OF VESICULAR STOMATITIS-VIRUS [J].
BLONDEL, D ;
HARMISON, GG ;
SCHUBERT, M .
JOURNAL OF VIROLOGY, 1990, 64 (04) :1716-1725
[4]   INTERACTIONS OF NORMAL AND MUTANT VESICULAR STOMATITIS-VIRUS MATRIX PROTEINS WITH THE PLASMA-MEMBRANE AND NUCLEOCAPSIDS [J].
CHONG, LD ;
ROSE, JK .
JOURNAL OF VIROLOGY, 1994, 68 (01) :441-447
[5]   MEMBRANE ASSOCIATION OF FUNCTIONAL VESICULAR STOMATITIS-VIRUS MATRIX PROTEIN INVIVO [J].
CHONG, LD ;
ROSE, JK .
JOURNAL OF VIROLOGY, 1993, 67 (01) :407-414
[6]   GENETIC-EVIDENCE FOR MULTIPLE FUNCTIONS OF THE MATRIX PROTEIN OF VESICULAR STOMATITIS-VIRUS [J].
COULON, P ;
DEUTSCH, V ;
LAFAY, F ;
MARTINETEDELIST, C ;
WYERS, F ;
HERMAN, RC ;
FLAMAND, A .
JOURNAL OF GENERAL VIROLOGY, 1990, 71 :991-996
[7]   Studies on the rabies virus RNA polymerase: 3. Two-dimensional electrophoretic analysis of the multiplicity of non-catalytic subunit (P protein) [J].
Eriguchi, Y ;
Toriumi, H ;
Kawai, A .
MICROBIOLOGY AND IMMUNOLOGY, 2002, 46 (07) :463-474
[8]   REVERSIBLE CONFORMATIONAL-CHANGES AND FUSION ACTIVITY OF RABIES VIRUS GLYCOPROTEIN [J].
GAUDIN, Y ;
TUFFEREAU, C ;
SEGRETAIN, D ;
KNOSSOW, M ;
FLAMAND, A .
JOURNAL OF VIROLOGY, 1991, 65 (09) :4853-4859
[9]   LOW-PH INDUCED CONFORMATIONAL-CHANGES IN VIRAL FUSION PROTEINS - IMPLICATIONS FOR THE FUSION MECHANISM [J].
GAUDIN, Y ;
RUIGROK, RWH ;
BRUNNER, J .
JOURNAL OF GENERAL VIROLOGY, 1995, 76 :1541-1556
[10]   FATTY ACYLATION OF RABIES VIRUS PROTEINS [J].
GAUDIN, Y ;
TUFFEREAU, C ;
BENMANSOUR, A ;
FLAMAND, A .
VIROLOGY, 1991, 184 (01) :441-444