Structure and function of disk aggregates of the coat protein of tobacco mosaic virus

Raghavendra, K.; Kelly, Judith A.; Khairallah, L. H.; Schuster, Todd M.

doi:10.1021/bi00420a002

Cited by 27 publications

(20 citation statements)

References 29 publications

(48 reference statements)

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“…After a 24-h equilibration at 20 "C, 62% by weight of TMVP sediments at 20.3 S and the remainder at 3.9 S. However, after a 24-h equilibration at 20 "C, 83% of the r-TMVP sediments at 28.6 S and 17% at 3.8 S. The r-TMVP sedimenting boundary of 28.6 S is as symmetrical as that seen for the 20s boundary of the wild-type protein, (Figure 4). This faster sedimenting species of r-TMVP is reminiscent of the 28s dimers of bilayer cylindrical disk aggregates formed by wild-type protein in high ionic strength buffers (Durham, 1972;Raghavendra et al, 1988). Figure 5 shows electron micrographs of aggregates of r-TMVP and TMVP from solutions at 20 "C and pH 7.0,6.0, and 5.5.…”

Section: Resultsmentioning

confidence: 93%

Preparation and properties of recombinant DNA derived tobacco mosaic virus coat protein

Shire¹,

McKay²,

Leung³

et al. 1990

Biochemistry

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Recombinant DNA derived tobacco mosaic virus (vulgare strain) coat protein (r-TMVP) was obtained by cloning and expression in Escherichia coli and was purified by column chromatography, self-assembly polymerization, and precipitation. SDS-PAGE, amino terminal sequencing, and immunoblotting with polyclonal antibodies raised against TMVP confirmed the identify and purity of the recombinant protein. Isoelectric focusing in 8 M urea and fast atom bombardment mass spectrometry demonstrated that the r-TMVP is not acetylated at the amino terminus, unlike the wild-type protein isolated from the tobacco plant derived virus. The characterization of r-TMVP with regard to its self-assembly properties revealed reversible endothermic polymerization as studied by analytical ultracentrifugation, circular dichroism, and electron microscopy. However, the details of the assembly process differed from those of the wild-type protein. At neutral pH, low ionic strength, and 20 degrees C, TMVP forms a 20S two-turn helical rod that acts as a nucleus for further assembly with RNA and additional TMVP to form TMV. Under more acidic conditions, this 20S structure also acts as a nucleus for protein self-assembly to form viruslike RNA-free rods. The r-TMVP that is not acetylated carries an extra positive charge at the amino terminus and does not appear to form the 20S nucleus. Instead, it forms a 28S four-layer structure, which resembles in size and structure the dimer of the bilayer disk formed by the wild-type protein at pH 8.0, high ionic strength, and 20 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS)

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Section: Resultsmentioning

confidence: 93%

Preparation and properties of recombinant DNA derived tobacco mosaic virus coat protein

Shire¹,

McKay²,

Leung³

et al. 1990

Biochemistry

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“…Moreover, it is noticeable that the S-value for the aggregate is given as 20.3S, while Durham (1972b) had shown that this rose abruptly from 19S to 420S with a small change of conditions and that the larger material could merge steadily into what were shown to be long helices. It is therefore quite probable that the aggregate(s) characterized by Schuster and his colleagues (Correia et al 1985;Raghavendra et al 1985Raghavendra et al , 1986Raghavendra et al , 1988 have indeed all been proto-helical, but not the same as the aggregate described as the`disk'. Moreover, a short proto-helix would not give the distinct two-layer appearance seen in side views of the disk in the electron microscope (see below).…”

Section: (B) Questions About the 20s Aggregate In Solutionmentioning

confidence: 99%

Self–assembly of tobacco mosaic virus: the role of an intermediate aggregate in generating both specificity and speed

Butler

1999

Phil. Trans. R. Soc. Lond. B

152

129

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The tobacco mosaic virus (TMV) particle was the ¢rst macromolecular structure to be shown to selfassemble in vitro, allowing detailed studies of the mechanism.Nucleation of TMV self-assembly is by the binding of a speci¢c stem-loop of the single-stranded viral RNA into the central hole of a two-ring sub-assembly of the coat protein, known as the`disk'. Binding of the loop onto its speci¢c binding site, between the two rings of the disk, leads to melting of the stem so more RNA is available to bind. The interaction of the RNA with the protein subunits in the disk cause this to dislocate into a proto-helix, rearranging the protein subunits in such a way that the axial gap between the rings at inner radii closes, entrapping the RNA.Assembly starts at an internal site on TMV RNA, about 1kb from its 3 H -terminus, and the elongation in the two directions is di¡erent. Elongation of the nucleated rods towards the 5 H -terminus occurs on à travelling loop' of the RNA and, predominantly, still uses the disk sub-assembly of protein subunits, consequently incorporating approximately 100 further nucleotides as each disk is added, while elongation towards the 3 H -terminus uses smaller protein aggregates and does not show this`quantized' incorporation.

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“…Whereas lipid rafts [1] are microdomains in the plasma membrane enriched in sphingolipids, and cholesterol, and implicated in the regulation of numerous signal transduction [2], membrane traffic pathways [3] and exocytosis [1], bilayered-disks composed exclusively of proteins have been studied in the context of protective coatings in viruses and virus reconstitution [4] and disks composed exclusively of micelle-forming surfactants (no lipids included in their composition) have been reported in a number of interesting studies [5][6][7][8][9][10][11][12]. This review is not about lipid rafts or protein bilayered disks (composed exclusively of proteins) or surfactant disks (composed exclusively of surfactants, no lipids included).…”

Section: Introductionmentioning

confidence: 99%

Lipid Bilayer Fragments and Disks in Drug Delivery

Carmona-Ribeiro

2006

CMC

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Certain lipids can be dispersed as colloidally stable bilayer fragments (BF) or disks with interesting properties for solubilization and delivery of hydrophobic or amphiphilic drugs. They were first observed and characterized as such in the nineties but remained silent in the literature regarding applications for drug delivery. Only recently their potential for delivery of hydrophobic drugs started to be realized. This review deals with electrostatically or sterically stabilized bilayer fragments which provided excellent solubilization sites for antifungal drugs, acted as drugs themselves against bacteria or fungus, could be loaded with amphiphilic drugs or produced lipid-covered drug particles to be delivered as a synergistic formulation in vivo.

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Structure and function of disk aggregates of the coat protein of tobacco mosaic virus

Cited by 27 publications

References 29 publications

Preparation and properties of recombinant DNA derived tobacco mosaic virus coat protein

Preparation and properties of recombinant DNA derived tobacco mosaic virus coat protein

Self–assembly of tobacco mosaic virus: the role of an intermediate aggregate in generating both specificity and speed

Lipid Bilayer Fragments and Disks in Drug Delivery

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