Crystallization and preliminary crystallographic analysis of the fusion core of the spike protein of the murine coronavirus mouse hepatitis virus (MHV)

被引:2
作者
Xu, YH
Bai, ZH
Qin, L
Li, X
Gao, G
Rao, ZH [1 ]
机构
[1] Tsing Hua Univ, Struct Biol Lab, Beijing 100084, Peoples R China
[2] Inst Biophys, Natl Lab Biomacromol, Beijing 100101, Peoples R China
[3] Univ Oxford, John Radcliffe Hosp, Nuffield Dept Clin Med, Oxford OX3 9DU, England
来源
ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY | 2004年 / 60卷
关键词
D O I
10.1107/S0907444904020517
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Crystals of a 2-Helix fusion-core construct of MHV spike protein (commonly referred to as E2) have been grown at 291 K using PEG 4000 as precipitant. The diffraction pattern of the crystal extends to 2.8 Angstrom resolution at 100 K in-house. Furthermore, a selenomethionine ( SeMet) derivative of MHV spike protein fusion core has been overexpressed and purified. The derivative crystals were obtained under similar conditions and three different wavelength data sets were collected to 2.4 Angstrom resolution from a single derivative crystal at BSRF (Beijing Synchrotron Radiation Facility). The crystals have unit-cell parameters a = b = 48.3, c = 199.6 Angstrom, alpha = beta = 90, gamma = 120degrees and belong to space group R3. Assuming the presence of two molecules in the asymmetric unit, the solvent content is calculated to be about 46%.
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页码:2013 / 2015
页数:3
相关论文
共 13 条
[1]   The coronavirus spike protein is a class I virus fusion protein: Structural and functional characterization of the fusion core complex [J].
Bosch, BJ ;
van der Zee, R ;
de Haan, CAM ;
Rottier, PJM .
JOURNAL OF VIROLOGY, 2003, 77 (16) :8801-8811
[2]   CORONAVIRUS IBV - STRUCTURAL CHARACTERIZATION OF THE SPIKE PROTEIN [J].
CAVANAGH, D .
JOURNAL OF GENERAL VIROLOGY, 1983, 64 (DEC) :2577-2583
[3]   MONOCLONAL-ANTIBODIES TO MURINE HEPATITIS VIRUS-4 (STRAIN-JHM) DEFINE THE VIRAL GLYCOPROTEIN RESPONSIBLE FOR ATTACHMENT AND CELL CELL-FUSION [J].
COLLINS, AR ;
KNOBLER, RL ;
POWELL, H ;
BUCHMEIER, MJ .
VIROLOGY, 1982, 119 (02) :358-371
[4]   EVIDENCE FOR A COILED-COIL STRUCTURE IN THE SPIKE PROTEINS OF CORONAVIRUSES [J].
DEGROOT, RJ ;
LUYTJES, W ;
HORZINEK, MC ;
VANDERZEIJST, BAM ;
SPAAN, WJM ;
LENSTRA, JA .
JOURNAL OF MOLECULAR BIOLOGY, 1987, 196 (04) :963-966
[5]   Mechanisms of viral membrane fusion and its inhibition [J].
Eckert, DM ;
Kim, PS .
ANNUAL REVIEW OF BIOCHEMISTRY, 2001, 70 :777-810
[6]   PROTEOLYTIC CLEAVAGE OF THE E2-GLYCOPROTEIN OF MURINE CORONAVIRUS - HOST-DEPENDENT DIFFERENCES IN PROTEOLYTIC CLEAVAGE AND CELL-FUSION [J].
FRANA, MF ;
BEHNKE, JN ;
STURMAN, LS ;
HOLMES, KV .
JOURNAL OF VIROLOGY, 1985, 56 (03) :912-920
[7]   THE COMPLETE SEQUENCE (22 KILOBASES) OF MURINE CORONAVIRUS GENE-1 ENCODING THE PUTATIVE PROTEASES AND RNA-POLYMERASE [J].
LEE, HJ ;
SHIEH, CK ;
GORBALENYA, AE ;
KOONIN, EV ;
LAMONICA, N ;
TULER, J ;
BAGDZHADZHYAN, A ;
LAI, MMC .
VIROLOGY, 1991, 180 (02) :567-582
[8]   PRIMARY STRUCTURE OF THE GLYCOPROTEIN-E2 OF CORONAVIRUS MHV-A59 AND IDENTIFICATION OF THE TRYPSIN CLEAVAGE SITE [J].
LUYTJES, W ;
STURMAN, LS ;
BREDENBEEK, PJ ;
CHARITE, J ;
VANDERZEIJST, BAM ;
HORZINEK, MC ;
SPAAN, WJM .
VIROLOGY, 1987, 161 (02) :479-487
[9]   Processing of X-ray diffraction data collected in oscillation mode [J].
Otwinowski, Z ;
Minor, W .
MACROMOLECULAR CRYSTALLOGRAPHY, PT A, 1997, 276 :307-326
[10]  
Siddell SG, 1995, The Coronaviridae, P1, DOI [DOI 10.1007/978-1-4899-1531-3_1, 10.1007/978-1-4899-1531-3_1]