A Synthetic Coiled-Coil Interactome Provides Heterospecific Modules for Molecular Engineering

Abstract

The versatile coiled-coil protein motif is widely used to induce and control macromolecular interactions in biology and materials science. Yet the types of interaction patterns that can be constructed using known coiled coils are limited. Here we greatly expand the coiled-coil toolkit by measuring the complete pair-wise interactions of 48 synthetic coiled coils and 7 human bZIP coiled coils using peptide microarrays. The resulting 55-member protein ‘interactome’ includes 27 pairs of interacting peptides that preferentially hetero-associate. The 27 pairs can be used in combinations to assemble sets of 3 to 6 proteins that compose networks of varying topologies. Of special interest are heterospecific peptide pairs that participate in mutually orthogonal interactions. Such pairs provide the opportunity to dimerize two separate molecular systems without undesired crosstalk. Solution and structural characterization of two such sets of orthogonal heterodimers provide details of their interaction geometries. The orthogonal pair, along with the many other network motifs discovered in our screen, provide new capabilities for synthetic biology and other applications.

Engineering robust control of two-component system phosphotransfer using modular scaffolds

“…Each scaffold additionally includes one of two different synthetic leucine zippers fused to the SH3 peptide with a nine-residue glycine/serine linker. Leucine zippers were chosen because of the large number of available high-affinity pairs (31,32) and their successful use in previous synthetic biology applications (6,33). Each scaffold leucine zipper (here termed "LZA" and "LZB") incorporated into the scaffold was specific for a corresponding zipper, LZa and LZb, that was fused to either CusR or CpxR, respectively, with a 10-or 12-residue glycine/serine linker.…”

High affinity receptor labeling based on basic leucine zipper domain peptides conjugated with pH-sensitive fluorescent dye: Visualization of AMPA-type glutamate receptor endocytosis in living neurons

“…The ZIP-tag is much smaller than other molecular tags, including antibodies, fluorescent proteins and αBTx. Heteromeric binding of ZIP peptides is based on 14 α-helical motifs that are wrapped around each other to form a super-helix, promoting tight packing of hydrophobic amino acid side chains (Reinke et al, 2010). The length of ZIP peptide complex is estimated to be 6.6 nm, similar to typical α-helical polypeptides.…”

A silk purse from a sow's ear—bioinspired materials based on α-helical coiled coils

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