bi0c00363_si_001.pdf (12.2 MB)
Enzyme-Directed Functionalization of Designed, Two-Dimensional Protein Lattices
journal contribution
posted on 2020-08-03, 16:33 authored by Rohit
H. Subramanian, Yuta Suzuki, Lorillee Tallorin, Swagat Sahu, Matthew Thompson, Nathan C. Gianneschi, Michael D. Burkart, F. Akif TezcanThe
design and construction of crystalline protein arrays to selectively
assemble ordered nanoscale materials have potential applications in
sensing, catalysis, and medicine. Whereas numerous designs have been
implemented for the bottom-up construction of protein assemblies,
the generation of artificial functional materials has been relatively
unexplored. Enzyme-directed post-translational modifications are responsible
for the functional diversity of the proteome and, thus, could be harnessed
to selectively modify artificial protein assemblies. In this study,
we describe the use of phosphopantetheinyl transferases (PPTases),
a class of enzymes that covalently modify proteins using coenzyme
A (CoA), to site-selectively tailor the surface of designed, two-dimensional
(2D) protein crystals. We demonstrate that a short peptide (ybbR)
or a molecular tag (CoA) can be covalently tethered to 2D arrays to
enable enzymatic functionalization using Sfp PPTase. The site-specific
modification of two different protein array platforms is facilitated
by PPTases to afford both small molecule- and protein-functionalized
surfaces with no loss of crystalline order. This work highlights the
potential for chemoenzymatic modification of large protein surfaces
toward the generation of sophisticated protein platforms reminiscent
of the complex landscape of cell surfaces.