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Structural analysis of membrane-bound retrovirus capsid proteins
被引:85
作者:
Barklis, E
McDermott, J
Wilkens, S
Schabtach, E
Schmid, MF
Fuller, S
Karanjia, S
Love, Z
Jones, R
Rui, YJ
Zhao, XM
Thompson, D
机构:
[1] OREGON HLTH SCI UNIV, DEPT MICROBIOL, PORTLAND, OR 97201 USA
[2] OREGON HLTH SCI UNIV, DEPT PATHOL, PORTLAND, OR 97201 USA
[3] UNIV OREGON, INST MOL BIOL, EUGENE, OR 97403 USA
[4] UNIV OREGON, INST NEUROSCI, EUGENE, OR 97403 USA
[5] BAYLOR COLL MED, VERNA & MARRS MCLEAN DEPT BIOCHEM, HOUSTON, TX 77030 USA
[6] BAYLOR COLL MED, WM KECK CTR COMPUTAT BIOL, HOUSTON, TX 77030 USA
[7] EUROPEAN MOL BIOL LAB, STRUCT BIOL PROGRAMME, D-69012 HEIDELBERG, GERMANY
[8] PURDUE UNIV, DEPT CHEM, W LAFAYETTE, IN 47907 USA
关键词:
electron microscopy;
image analysis;
lipid;
monolayer;
retrovirus;
D O I:
10.1093/emboj/16.6.1199
中图分类号:
Q5 [生物化学];
Q7 [分子生物学];
学科分类号:
071010 ;
081704 ;
摘要:
We have developed a system for analysis of histidine-tagged (His-tagged) retrovirus core (Gag) proteins, assembled in vitro on lipid monolayers consisting of egg phosphatidylcholine (PC) plus the novel lipid DHGN, DHGN was shown to chelate nickel by atomic absorption spectrometry, and DHGN-containing monolayers specifically bound gold conjugates of His-tagged proteins, Using PC+DHGN monolayers, we examined membrane-bound arrays of an N-terminal His-tagged Moloney murine leukemia virus (M-MuLV) capsid (CA) protein, His-MoCA, and in vivo studies suggest that in vitro-derived His-MoCA arrays reflect some of the Gag protein interactions which occur in assembling virus particles, The His-MoCA proteins formed extensive two-dimensional (2D) protein crystals, with reflections out to 9.5 Angstrom resolution, The image-analyzed 2D projection of His-MoCA arrays revealed a distinct cage-like network, The asymmetry of the individual building blocks of the network led to the formation of two types of hexamer rings, surrounding protein-free cage holes, These results predict that Gag hexamers constitute a retrovirus core substructure, and that cage hole sizes define an exclusion limit for entry of retrovirus envelope proteins, or other plasma membrane proteins, into virus particles, We believe that the 2D crystallization method will permit the detailed analysis of retroviral Gag proteins and other His-tagged proteins.
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页码:1199 / 1213
页数:15
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