sb7b00447_si_001.pdf (998.4 kB)
Programmed Protein Self-Assembly Driven by Genetically Encoded Intein-Mediated Native Chemical Ligation
journal contribution
posted on 2018-02-23, 00:00 authored by Joseph
A. Harvey, Laura S. Itzhaki, Ewan R. G. MainHarnessing
and controlling self-assembly is an important step in
developing proteins as novel biomaterials. With this goal, here we
report the design of a general genetically programmed system that
covalently concatenates multiple distinct protein domains into specific
assembled arrays. It is driven by iterative intein-mediated native
chemical ligation (NCL) under mild native conditions. The system uses
a series of initially inert recombinant protein fusions that sandwich
the protein modules to be ligated between one of a number of different
affinity tags and an intein protein domain. Orthogonal activation
at opposite termini of compatible protein fusions, via protease and
intein cleavage, coupled with sequential mixing directs an irreversible
and traceless stepwise assembly process. This gives total control
over the composition and arrangement of component proteins within
the final product, enabled the limits of the systemreaction
efficiency and yieldto be investigated, and led to the production
of “functional” assemblies.