As a prototype of “click” chemistry, the Cu(I) catalyzed azide-alkyne cycloaddition (CuAAC) reaction has flourished with applications in organic synthesis, polymer and material science, and biotechnology. In particular, this reaction has shown great promise as a secondary conjugation method for proteins and bionanoparticles. Thus far, azide and alkyne incorporation has been accomplished using traditional bioconjugation methods targeting lysine, cysteine and acidic amino acids. Recently tyrosine residues have been targeted with a diazonium-coupling reaction, which can efficiently target the phenolic group of tyrosine residues at physiological conditions. This reaction is very efficient, yet has two distinct disadvantages for broader applications. First, synthesis of desired starting materials is difficult; and second, the reaction is only suitable for electron-deficient anilines. Here we demonstrate that terminal alkyne groups can be attached to tyrosine residues and the sequential CuAAC reaction can efficiently be used for the bioconjugation of plant virus, tobacco mosaic virus (TMV). TMV was chosen as a rod-like (300 nm in length) multivalent protein scaffold to demonstrate this strategy. The quantitative covalent attachment of polymers, peptides and small molecules is shown using this combination of methods. To highlight the modularity and robustness of this CuAAC conjugation, the cell adhesion was studied after TMV was immobilized on a glass surface with a variety of groups that can promote or prevent cell binding.