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This paper presents a comprehensive study of the self-assembly dynamics and the biosensing efficacy of Tobacco mosaic virus-like particle (TMV VLP) sensing probes using an impedimetric microsensor platform. TMV VLPs are high surface area macromolecules with nanorod structures constructed from helical arrangements of thousands of identical coat proteins. Genetically modified TMV VLPs express both surface attachment-promoting cysteine residues and FLAG-tag antibody binding peptides on their coat protein outer surfaces, making them selective biosensing probes with self-assembly capability on sensors. The VLP self-assembly dynamics were studied by the continuous monitoring of impedance changes at 100Hz using interdigitated impedimetric microsensors. Electrical impedance spectroscopy revealed VLP saturation on impedance sensor surface with the coverage of 68% in self-assembly process. The VLP-functionalized impedance sensors responded to 12ng/ml to 1.2μg/ml of target anti-FLAG IgG antibodies in the subsequent enzyme-linked immunosorbent assays (ELISA), and yielded 18-35% total impedance increases, respectively. The detection limit of the target antibody is 9.1ng/ml using the VLP-based impedimetric microsensor. These results highlight the significant potential of genetically modified VLPs as selective nanostructured probes for autonomous sensor functionalization and enhanced biosensing., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Uniform dimensions and genetic tractability make filamentous viruses attractive templates for the display of functional groups used in materials science, sensor applications, and vaccine development. However, active virus replication and recombination often limit the usefulness of these viruses for such applications. To circumvent these limitations, genetic modifications of selected negatively charged intersubunit carboxylate residues within the coat protein of tobacco mosaic virus (TMV) were neutralized so as to stabilize the assembly of rod-shaped virus-like particles (VLPs) within bacterial expression systems. Here we show that TMV-VLP nanorods are easily purified, stable, and can be programmed in a variety of configurations to display functional peptides for antibody and small molecule binding.

Designed as a high-capacity alloy host for Na-ion chemistry, a forest of Sn nanorods with a unique core-shell structure was synthesized on viral scaffolds, which were genetically engineered to ensure a nearly vertical alignment upon self-assembly onto a metal substrate. The interdigital spaces thus formed between individual rods effectively accommodated the volume expansion and contraction of the alloy upon sodiation/desodiation, while additional carbon-coating engineered over these nanorods further suppressed Sn aggregation during extended electrochemical cycling. Due to the unique nanohierarchy of multiple functional layers, the resultant 3D nanoforest of C/Sn/Ni/TMV1cys, binder-free composite electrode already and evenly assembled on a stainless steel current collector, exhibited supreme capacity utilization and cycling stability toward Na-ion storage and release. An initial capacity of 722 mA·h (g Sn)(-1) along with 405 mA·h (g Sn)(-1) retained after 150 deep cycles demonstrates the longest-cycling nano-Sn anode material for Na-ion batteries reported in the literature to date and marks a significant performance improvement for neat Sn material as alloy host for Na-ion chemistry.

The structural proteins of plant viruses have evolved to self-associate into complex macromolecules that are centrally involved in virus biology. In this review, the structural and biophysical properties of the Tobacco mosaic virus (TMV) coat protein (CP) are addressed in relation to its role in host resistance and disease development. TMV CP affects the display of several specific virus and host responses, including cross-protection, systemic virus movement, hypersensitive disease resistance, and symptom development. Studies indicate that the three-dimensional structure of CP is critical to the control of these responses, either directly through specific structural motifs or indirectly via alterations in CP assembly. Thus, both the structure and assembly of the TMV CP function as determinants in the induction of disease and resistance responses.

The hypersensitivity resistance response directed by the N' gene in Nicotiana sylvestris is elicited by the tobacco mosaic virus (TMV) coat protein R46G, but not by the U1 wild-type TMV coat protein. In this study, the structural and hydrodynamic properties of R46G and wild-type coat proteins were compared for variations that may explain N' gene elicitation. Circular dichroism spectroscopy reveals no significant secondary or tertiary structural differences between the elicitor and nonelicitor coat proteins. Analytical ultracentrifugation studies, however, do show different concentration dependencies of the weight average sedimentation coefficients at 4 degrees C. Viral reconstitution kinetics at 20 degrees C were used to determine viral assembly rates and as an initial assay of the rate of 20S formation, the obligate species for viral reconstitution. These kinetic results reveal a decreased lag time for reconstitution performed with R46G that initially lack the 20S aggregate. However, experiments performed with 20S initially present reveal no detectable differences indicating that the mechanism of viral assembly is similar for the two coat protein species. Therefore, an increased rate of 20S formation from R46G subunits may explain the differences in the viral reconstitution lag times. The inferred increase in the rate of 20S formation is verified by direct measurement of the 20S boundary as a function of time at 20 degrees C using velocity sedimentation analysis. These results are consistent with the interpretation that there may be an altered size distribution and/or lifetime of the small coat protein aggregates in elicitors that allows N. sylvestris to recognize the invading virus.

To investigate the molecular role of the tobacco mosaic tobamovirus (TMV) coat protein (CP) in conferring cross-protection, a potato X potexvirus (PVX) vector (S. Chapman, Plant J. 2, 549-557, 1992) was used to systemically express a set of TMV mutant CPs in Nicotiana benthamiana prior to challenge inoculation with TMV. PVX-expressed wild-type TMV CP delayed TMV accumulation for up to 2 weeks compared to unprotected plants or plants preinfected with the unmodified PVX vector. Similar delays in TMV accumulation were obtained using TMV CPs that were deficient in virion formation but competent to assemble into helical aggregates. In contrast, TMV CPs that were incapable of helical aggregation or unable to bind viral RNA did not delay the accumulation of TMV. Furthermore, TMV CPs with enhanced intersubunit interactions that favor helical aggregation produced significantly greater delays in the accumulation of challenge TMV than obtained from the wild-type CP. Thus the capabilities of TMV CP to interact with viral RNA and self-associate in a helical fashion appear to be essential to its ability to confer protection. Taken together, these findings support a model for CP-mediated resistance in which the protecting CP recoats the challenge virus RNA as it disassembles., (Copyright 1998 Academic Press.)