Structural Analysis of Membrane-bound Retrovirus Capsid Proteins

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 1997 Mar 17

PMID 9135137

Citations 33

Authors

E Barklis

J McDermott

S Wilkens

E Schabtach

M F Schmid

S Fuller

S Karanjia

Z Love

R Jones

Y Rui

X Zhao

D Thompson

Affiliations

Soon will be listed here.

Abstract

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 A 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.

Citing Articles

Capsid-specific nanobody effects on HIV-1 assembly and infectivity.

Alfadhli A, Romanaggi C, Barklis R, Merutka I, Bates T, Tafesse F Virology. 2021; 562:19-28.

PMID: 34246112 PMC: 8419096. DOI: 10.1016/j.virol.2021.07.001.

Affinity Capture of p97 with Small-Molecule Ligand Bait Reveals a 3.6 Å Double-Hexamer Cryoelectron Microscopy Structure.

Hoq M, Vago F, Li K, Kovaliov M, Nicholas R, Huryn D ACS Nano. 2021; 15(5):8376-8385.

PMID: 33900731 PMC: 11752781. DOI: 10.1021/acsnano.0c10185.

Organization of Farnesylated, Carboxymethylated KRAS4B on Membranes.

Barklis E, Stephen A, Staubus A, Barklis R, Alfadhli A J Mol Biol. 2019; 431(19):3706-3717.

PMID: 31330153 PMC: 6733658. DOI: 10.1016/j.jmb.2019.07.025.

Selective Capture of Histidine-tagged Proteins from Cell Lysates Using TEM grids Modified with NTA-Graphene Oxide.

Benjamin C, Wright K, Bolton S, Hyun S, Krynski K, Grover M Sci Rep. 2016; 6:32500.

PMID: 27748364 PMC: 5066248. DOI: 10.1038/srep32500.

Nonfouling NTA-PEG-Based TEM Grid Coatings for Selective Capture of Histidine-Tagged Protein Targets from Cell Lysates.

Benjamin C, Wright K, Hyun S, Krynski K, Yu G, Bajaj R Langmuir. 2016; 32(2):551-9.

PMID: 26726866 PMC: 5310270. DOI: 10.1021/acs.langmuir.5b03445.

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