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Download fileBioorthogonal Modification of the Major Sheath Protein of Bacteriophage M13: Extending the Versatility of Bionanomaterial Scaffolds
journal contribution
posted on 2016-09-14, 00:00 authored by Taylor Urquhart, Elisabeth Daub, John Frank HonekWith a mass of ∼1.6 ×
107 Daltons and composed
of approximately 2700 proteins, bacteriophage M13 has been employed
as a molecular scaffold in bionanomaterials fabrication. In order
to extend the versatility of M13 in this area, residue-specific unnatural
amino acid incorporation was employed to successfully display azide
functionalities on specific solvent-exposed positions of the pVIII
major sheath protein of this bacteriophage. Employing a combination
of engineered mutants of the gene coding for the pVIII protein, the
methionine (Met) analog, l-azidohomoalanine (Aha), and a
suitable Escherichia coli Met auxotroph
for phage production, conditions were developed to produce M13 bacteriophage
labeled with over 350 active azides (estimated by fluorescent dye
labeling utilizing a strain-promoted azide–alkyne cycloaddition)
and capable of azide-selective attachment to 5 nm gold nanoparticles
as visualized by transmission electron microscopy. The capability
of this system to undergo dual labeling utilizing both chemical acylation
and bioorthogonal cycloaddition reactions was also verified. The above
stratagem should prove particularly advantageous in the preparation
of assemblies of larger and more complex molecular architectures based
on the M13 building block.
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Keywords
acid incorporationMajor Sheath ProteinBioorthogonal Modificationbioorthogonal cycloaddition reactionsM 13 building blockpVIII proteinchemical acylationBionanomaterial ScaffoldsM 13solvent-exposed positionstransmission electron microscopybionanomaterials fabricationsheath proteinphage productionM 13 bacteriophageEscherichia coli Met auxotroph5 nm gold nanoparticlesbacteriophage M 132700 proteinsazide-selective attachmentdisplay azide functionalitiesgene coding