Chemoselective Modification of Turnip Yellow Mosaic Virus by Cu(I) Catalyzed Azide−Alkyne 1,3-Dipolar Cycloaddition Reaction and Its Application in Cell Binding

Abstract

Turnip yellow mosaic virus (TYMV) is an icosahedral plant virus with a diameter of 28-30 nm that can be isolated in gram quantities from turnip or Chinese cabbage inexpensively. In this study, TYMV combined with spatially addressable surface chemistries was selected as a prototype bionanoparticle for modulating patterns of cell adhesion, morphology, and proliferation. We exploited the chemical reactivity of TYMV using the mild conditions of Cu(I) catalyzed azide-alkyne cycloaddition (CuAAC) reaction, the best example of "click" chemistry. Oligo-ethylene glycol (OEG) short chain, coumarintriazole, and RGD-containing peptide were grafted on the surface of TYMV via carbodiimide activation and CuAAC reaction. The bioconjugation to intact viral particles was confirmed by MS, TEM, FPLC, and SDS-PAGE with fluorescence visualization analysis. Therefore, this method is a generally useful means of incorporating various types of functionalities onto the TYMV surface. Further studies were done to learn the behavior of NIH-3T3 fibroblast cells on the modified or unmodified TYMV surfaces. OEG-modified TYMV surfaces retarded cell attachment and growth, while cell adhesion, spreading, and proliferation were dramatically enhanced on RGD-modified TYMV surfaces. Compared with RGD immobilized 3-aminopropyltriethoxysilane-coated glass surface, the cells are more ready to spread fully and proliferate on TYMV-RGD coated surface, which thus provides a more cell-friendly environment with nanometer-scale surface features. This illustrates the potential application of plant virus based materials in tissue engineering, drug delivery, and biosensing.

Surface modification of cowpea chlorotic mottle virus capsids via a copper(I)-catalyzed azide-alkyne cycloaddition (CuAAC) reaction and their adhesion behavior with HeLa cells

“…Additionally, the RGD ligand density and its spatial arrangement could play a significant role during the binding process [36]. Another difference from previously reported results [21,28] was that the HeLa cells did not present similar elongated or stretched morphologies to those of NIH3T3 fibroblasts on APTES slides. This might be attributable to the different properties of these two cell types and their interactions with the APTES ligand on the slides.…”

“…The reactivity of the CCMV carboxyl groups was tested with fluorescein cadaverine under activation with EDC and sulfo-NHS, which revealed that 560 carboxylates were modified in high yields [12]. This was approximately 4.7 6 .2 times higher than that accessible in TYMV (90 ~ 120 carboxylic groups) [21]. To explore the potential of introducing a multifunctional group onto the surface of CCMV, the viral capsids were labeled with acetylene at surface-exposed carboxylic groups.…”

“…The 3-aminopropyltriethoxysilane (APTES)-functionalized slides were purchased from Labscientific. The CCMV capsids were prepared as described in the literature [21]. The purified CCMV sample (1 mg/mL) in 80% assembly buffer (100 mM sodium acetate, 1 mM EDTA, 5 mM DTT, 0.5 mM PMSF, and pH 5.2) plus 20% dimethyl sulfoxide (DMSO) was used for the process.…”

“…The tyrosine residues on the helical surface of tobacco mosaic virus (TMV) were coupled with chemoselective diazonium and azides via a sequential CuAAC reaction [20]. Various fluorescent coumarins and the Arg-Gly-Asp (RGD) peptide have also been grafted onto the surface of the turnip yellow mosaic virus (TYMV) by the CuAAC reaction [21].…”

Plant virus directed fabrication of nanoscale materials and devices

“…Formation of PVN films by these methods is predominantly controlled by the concentration of PVN and the strength of its surface attraction (Barick and Bahadur, 2010;Evans, 2008). The uniform and multivalent nature of these PVN coated surfaces has led to investigation into their effects on cell responses and differentiation (Kaur et al, 2010a(Kaur et al, , 2010bZan et al, 2012;Zeng et al, 2011). Using Turnip yellow mosaic virus (TYMV) as a scaffold for the multivalent display of the cell adhesion peptide RGD, Zan et al demonstrated enhanced mesenchymal stem cell adhesion and spreading onto surfaces layered with TYMV genetically programed to display the RGD peptide (Zan et al, 2012).…”