Assembly of Multilayer Arrays of Viral Nanoparticles via Biospecific Recognition: A Quartz Crystal Microbalance with Dissipation Monitoring Study

Steinmetz, Nicole F.; Bock, E.; Richter, Ralf P.; Spatz, Joachim P.; Lomonossoff, George P.; Evans, David

doi:10.1021/bm700797b

Cited by 55 publications

(56 citation statements)

References 42 publications

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“…Grafting was again mediated by a biotin tag. A linker of about 3 nm in contour length (47) was placed between the biotin and the reducing end of HA to enhance the conformational freedom of the surface-bound oHA chains. Each oHA chain is 4.5 nm long and can interact with only one TSG-6 molecule at a time, and previous studies have shown that ϳ5 sugar monomers probably fill its HA-binding site (48).…”

Section: Design Of Ha Model Films To Study Tsg-6-hyaluronanmentioning

confidence: 99%

The Inflammation-associated Protein TSG-6 Cross-links Hyaluronan via Hyaluronan-induced TSG-6 Oligomers

Baranova

Nilebäck

Haller³

et al. 2011

Journal of Biological Chemistry

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131

190

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Tumor necrosis factor-stimulated gene-6 (TSG-6) is a hyaluronan (HA)-binding protein that plays important roles in inflammation and ovulation. TSG-6-mediated cross-linking of HA has been proposed as a functional mechanism (e.g. for regulating leukocyte adhesion), but direct evidence for cross-linking is lacking, and we know very little about its impact on HA ultrastructure. Here we used films of polymeric and oligomeric HA chains, end-grafted to a solid support, and a combination of surface-sensitive biophysical techniques to quantify the binding of TSG-6 into HA films and to correlate binding to morphological changes. We find that full-length TSG-6 binds with pronounced positive cooperativity and demonstrate that it can cross-link HA at physiologically relevant concentrations. Our data indicate that cooperative binding of full-length TSG-6 arises from HA-induced protein oligomerization and that the TSG-6 oligomers act as cross-linkers. In contrast, the HA-binding domain of TSG-6 (the Link module) alone binds without positive cooperativity and weaker than the full-length protein.Both the Link module and full-length TSG-6 condensed and rigidified HA films, and the degree of condensation scaled with the affinity between the TSG-6 constructs and HA. We propose that condensation is the result of protein-mediated HA crosslinking. Our findings firmly establish that TSG-6 is a potent HA cross-linking agent and might hence have important implications for the mechanistic understanding of the biological function of TSG-6 (e.g. in inflammation). Hyaluronan (HA)3 is a structurally simple and linear polysaccharide. It is ubiquitous in the extracellular matrix of vertebrates and plays important roles in numerous physiological and pathological processes, such as inflammation, fertilization, embryogenesis, tumor development, osteoarthritis, and atherosclerosis (1, 2). HA is considered a "pericellular cue" (3) (i.e. it serves as a versatile scaffold within which other molecules are organized and regulated). A number of proteins, called hyaladherins (4), can bind to the flexible HA chains and engender self-assembly into large and hydrated multimolecular complexes (5-7).The secreted product of tumor necrosis factor-stimulated gene-6 (TSG-6) (8, 9) is of particular importance for the formation and remodeling of HA-rich pericellular coats (10, 11) and extracellular matrices (12). There is little or no constitutive expression of TSG-6 in most adult tissues (with the exception of bone marrow (13) and epidermis (14)). Expression is elevated in response to stimulation with proinflammatory mediators or certain growth factors (8, 9, 15-18), and TSG-6 is detected in the context of many inflammatory diseases (19,20) and in inflammation-like processes, such as ovulation (21,22). TSG-6 is composed mainly of two contiguous domains, a Link module and a CUB module (8,17,23,24). The Link module is conserved among members of the hyaladherin family (4) and is essential for binding to HA (23). Administration of recombinant human TSG-6 Link module (L...

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Section: Design Of Ha Model Films To Study Tsg-6-hyaluronanmentioning

confidence: 99%

The Inflammation-associated Protein TSG-6 Cross-links Hyaluronan via Hyaluronan-induced TSG-6 Oligomers

Baranova

Nilebäck

Haller³

et al. 2011

Journal of Biological Chemistry

Self Cite

131

190

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“…9). To study the properties of the multilayers, CPMV LbL assembly was followed in real time and in situ using quartz crystal microbalance with dissipation monitoring (QCMD; Steinmetz et al 2008a). In particular, the effect of modifying the density of biotin molecules attached via longer and shorter linkers to the virus surface was investigated.…”

Section: Cowpea Mosaic Virusmentioning

confidence: 99%

Applications of Plant Viruses in Bionanotechnology

Lomonossoff

Evans

2011

Current Topics in Microbiology and Immunology

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The capsids of most plant viruses are simple and robust structures consisting of multiple copies of one or a few types of protein subunit arranged with either icosahedral or helical symmetry. In many cases, capsids can be produced in large quantities either by the infection of plants or by the expression of the subunit(s) in a variety of heterologous systems. In view of their relative simplicity, stability and ease of production, plant virus particles or virus-like particles (VLPs) have attracted attention as potential reagents for applications in bionanotechnology. As a result, plant virus particles have been subjected to both genetic and chemical modification, have been used to encapsulate foreign material and have, themselves, been incorporated into supramolecular structures.

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“…The merged images confirmed that the virions sat one on top of the other; this is in contrast with a mixed-layer monolayer, where the images do not line up, as the particles are occupying the same layer and competing for binding sites. QCMD (quartz crystal microbalance with dissipation monitoring) studies of the alternating deposition of biotin-modified CPMV, with different biotin-spacer length, and SAv on a biotinylated solid-supported lipid bilayer showed that the level of biotin coverage and the length of the biotin spacer affect the degree of intra-and inter-layer cross-linking [15]. A long spacer and high biotin coverage gave the most regular and densest arrays; the degree of layer coverage and of cross-linking can therefore be highly tuned.…”

Section: Layer and Array Formationmentioning

confidence: 99%

Exploitation of plant and archaeal viruses in bionanotechnology

Evans

2009

Biochemical Society Transactions

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CPMV (cowpea mosaic virus), a plant virus, is a naturally occurring sphere-like nanoparticle, and is used as a synthon and/or template in bionanoscience. The virions formed by CPMV can be regarded as programmable nanobuilding blocks with a diameter of approximately 30 nm. A range of molecules have been attached to this viral nanoscaffold, yielding stable nanoparticles that display multiple copies of the desired molecule. It has been shown that, in addition to surface amine groups, surface carboxy groups are also addressable, and a procedure has been developed that enables introduction of reactive thiols at the virion surface that avoids virus aggregation. Furthermore, the virions can be functionalized to form electroactive nanoparticles. Methods for the construction of arrays and multilayers, using a layer-by-layer approach, have been established. As proof of concept, for example, CPMV particles have been immobilized on surfaces and arranged in defined layers. Engineered variants of CPMV can be used as templates for mineralization with, for example, silica to give monodisperse robust silica nanoparticles of approximately 32 nm. SIRV2 (Sulfolobus islandicus rod-shaped virus 2), is a robust archaeal virus, resistant to high temperature and low pH. SIRV2 can act as a template for site-selective and spatially controlled chemical modification. Both the ends and the body of the virus, or the ends only, can be chemically addressed; SIRV2 can be regarded as a structurally unique nanobuilding block.

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Assembly of Multilayer Arrays of Viral Nanoparticles via Biospecific Recognition: A Quartz Crystal Microbalance with Dissipation Monitoring Study

Cited by 55 publications

References 42 publications

The Inflammation-associated Protein TSG-6 Cross-links Hyaluronan via Hyaluronan-induced TSG-6 Oligomers

The Inflammation-associated Protein TSG-6 Cross-links Hyaluronan via Hyaluronan-induced TSG-6 Oligomers

Applications of Plant Viruses in Bionanotechnology

Exploitation of plant and archaeal viruses in bionanotechnology

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