16 research outputs found
Design Strategies for the Sequence-Based Mimicry of Side-Chain Display in Protein β‑Sheets by α/β-Peptides
The sophistication of folding patterns and functions
displayed
by unnatural-backbone oligomers has increased tremendously in recent
years. Design strategies for the mimicry of tertiary structures seem
within reach; however, a general method for the mimicry of sheet segments
in the context of a folded protein is an unmet need preventing realization
of this goal. Previous work has shown that 1→1 α→β-residue
substitutions at cross-strand positions in a hairpin-forming α-peptide
sequence can generate an α/β-peptide analogue that folds
in aqueous conditions but with a change in side-chain display relative
to the natural sequence; this change would prevent application of
single β-residue substitutions in a larger protein. Here, we
evaluate four different substitution strategies based on replacement
of αα dipeptide segments for the ability to retain both
sheet folding encoded by a parent α-peptide sequence as well
as nativelike side-chain display in the vicinity of the β-residue
insertion point. High-resolution structure determination and thermodynamic
analysis of folding by multidimensional NMR suggest that three of
the four designs examined are applicable to larger proteins
Design Strategies for the Sequence-Based Mimicry of Side-Chain Display in Protein β‑Sheets by α/β-Peptides
The sophistication of folding patterns and functions
displayed
by unnatural-backbone oligomers has increased tremendously in recent
years. Design strategies for the mimicry of tertiary structures seem
within reach; however, a general method for the mimicry of sheet segments
in the context of a folded protein is an unmet need preventing realization
of this goal. Previous work has shown that 1→1 α→β-residue
substitutions at cross-strand positions in a hairpin-forming α-peptide
sequence can generate an α/β-peptide analogue that folds
in aqueous conditions but with a change in side-chain display relative
to the natural sequence; this change would prevent application of
single β-residue substitutions in a larger protein. Here, we
evaluate four different substitution strategies based on replacement
of αα dipeptide segments for the ability to retain both
sheet folding encoded by a parent α-peptide sequence as well
as nativelike side-chain display in the vicinity of the β-residue
insertion point. High-resolution structure determination and thermodynamic
analysis of folding by multidimensional NMR suggest that three of
the four designs examined are applicable to larger proteins
Design Strategies for the Sequence-Based Mimicry of Side-Chain Display in Protein β‑Sheets by α/β-Peptides
The sophistication of folding patterns and functions
displayed
by unnatural-backbone oligomers has increased tremendously in recent
years. Design strategies for the mimicry of tertiary structures seem
within reach; however, a general method for the mimicry of sheet segments
in the context of a folded protein is an unmet need preventing realization
of this goal. Previous work has shown that 1→1 α→β-residue
substitutions at cross-strand positions in a hairpin-forming α-peptide
sequence can generate an α/β-peptide analogue that folds
in aqueous conditions but with a change in side-chain display relative
to the natural sequence; this change would prevent application of
single β-residue substitutions in a larger protein. Here, we
evaluate four different substitution strategies based on replacement
of αα dipeptide segments for the ability to retain both
sheet folding encoded by a parent α-peptide sequence as well
as nativelike side-chain display in the vicinity of the β-residue
insertion point. High-resolution structure determination and thermodynamic
analysis of folding by multidimensional NMR suggest that three of
the four designs examined are applicable to larger proteins
Tuning Assembly Size in Peptide-Based Supramolecular Polymers by Modulation of Subunit Association Affinity
Nature
uses proteins and nucleic acids to form a wide array of
functional architectures, and scientists have found inspiration from
these structures in the rational design of synthetic biomaterials.
We have recently shown that a modular subunit consisting of two α-helical
coiled coil peptides attached at their midpoints by an organic linking
group can spontaneously self-assemble in aqueous solution to form
a soluble supramolecular polymer. Here we explore the use of coiled-coil
association affinity, readily tuned by amino acid sequence, as a means
to predictably alter properties of these supramolecular assemblies.
A series of dimeric coiled-coil peptide sequences with identical quaternary
folded structures but systematically altered folded stability were
designed and biophysically characterized. The sequences were cross-linked
to generate a series of branched, self-assembling biomacromolecular
subunits. A clear relationship is observed between coiled-coil association
affinity and apparent hydrodynamic diameter of the supramolecular
polymers formed by these subunits. Our results provide a family of
soluble supramolecular polymers of tunable size and well-characterized
coiled-coil sequences that add to the library of building blocks available
for use in the rational design of protein-based supramolecular biomaterials
Design Strategies for the Sequence-Based Mimicry of Side-Chain Display in Protein β‑Sheets by α/β-Peptides
The sophistication of folding patterns and functions
displayed
by unnatural-backbone oligomers has increased tremendously in recent
years. Design strategies for the mimicry of tertiary structures seem
within reach; however, a general method for the mimicry of sheet segments
in the context of a folded protein is an unmet need preventing realization
of this goal. Previous work has shown that 1→1 α→β-residue
substitutions at cross-strand positions in a hairpin-forming α-peptide
sequence can generate an α/β-peptide analogue that folds
in aqueous conditions but with a change in side-chain display relative
to the natural sequence; this change would prevent application of
single β-residue substitutions in a larger protein. Here, we
evaluate four different substitution strategies based on replacement
of αα dipeptide segments for the ability to retain both
sheet folding encoded by a parent α-peptide sequence as well
as nativelike side-chain display in the vicinity of the β-residue
insertion point. High-resolution structure determination and thermodynamic
analysis of folding by multidimensional NMR suggest that three of
the four designs examined are applicable to larger proteins
Design Strategies for the Sequence-Based Mimicry of Side-Chain Display in Protein β‑Sheets by α/β-Peptides
The sophistication of folding patterns and functions
displayed
by unnatural-backbone oligomers has increased tremendously in recent
years. Design strategies for the mimicry of tertiary structures seem
within reach; however, a general method for the mimicry of sheet segments
in the context of a folded protein is an unmet need preventing realization
of this goal. Previous work has shown that 1→1 α→β-residue
substitutions at cross-strand positions in a hairpin-forming α-peptide
sequence can generate an α/β-peptide analogue that folds
in aqueous conditions but with a change in side-chain display relative
to the natural sequence; this change would prevent application of
single β-residue substitutions in a larger protein. Here, we
evaluate four different substitution strategies based on replacement
of αα dipeptide segments for the ability to retain both
sheet folding encoded by a parent α-peptide sequence as well
as nativelike side-chain display in the vicinity of the β-residue
insertion point. High-resolution structure determination and thermodynamic
analysis of folding by multidimensional NMR suggest that three of
the four designs examined are applicable to larger proteins
Heterogeneous-Backbone Foldamer Mimics of Zinc Finger Tertiary Structure
A variety of oligomeric
backbones with compositions deviating from
biomacromolecules can fold in defined ways. Termed “foldamers,”
these agents have diverse potential applications. A number of protein-inspired
secondary structures (e.g., helices, sheets) have been produced from
unnatural backbones, yet examples of tertiary folds combining several
secondary structural elements in a single entity are rare. One promising
strategy to address this challenge is the systematic backbone alteration
of natural protein sequences, through which a subset of the native
side chains is displayed on an unnatural building block to generate
a heterogeneous backbone. A drawback to this approach is that substitution
at more than one or two sites often comes at a significant energetic
cost to fold stability. Here we report heterogeneous-backbone foldamers
that mimic the zinc finger domain, a ubiquitous and biologically important
metal-binding tertiary motif, and do so with a folded stability that
is superior to the natural protein on which their design is based.
A combination of UV–vis spectroscopy, isothermal titration
calorimetry, and multidimensional NMR reveals that suitably designed
oligomers with >20% modified backbones can form native-like tertiary
folds with metal-binding environments identical to the prototype sequence
(the third finger of specificity factor 1) and enhanced thermodynamic
stability. These results expand the scope of heterogeneous-backbone
foldamer design to a new tertiary structure class and show that judiciously
applied backbone modification can be accompanied by improvement to
fold stability
Design Strategies for the Sequence-Based Mimicry of Side-Chain Display in Protein β‑Sheets by α/β-Peptides
The sophistication of folding patterns and functions
displayed
by unnatural-backbone oligomers has increased tremendously in recent
years. Design strategies for the mimicry of tertiary structures seem
within reach; however, a general method for the mimicry of sheet segments
in the context of a folded protein is an unmet need preventing realization
of this goal. Previous work has shown that 1→1 α→β-residue
substitutions at cross-strand positions in a hairpin-forming α-peptide
sequence can generate an α/β-peptide analogue that folds
in aqueous conditions but with a change in side-chain display relative
to the natural sequence; this change would prevent application of
single β-residue substitutions in a larger protein. Here, we
evaluate four different substitution strategies based on replacement
of αα dipeptide segments for the ability to retain both
sheet folding encoded by a parent α-peptide sequence as well
as nativelike side-chain display in the vicinity of the β-residue
insertion point. High-resolution structure determination and thermodynamic
analysis of folding by multidimensional NMR suggest that three of
the four designs examined are applicable to larger proteins
Protein-like Tertiary Folding Behavior from Heterogeneous Backbones
Because proteins play vital roles
in life, much effort has been
invested in their mimicry by synthetic agents. One approach is to
design unnatural backbone oligomers (“foldamers”) that
fold like natural peptides. Despite success in secondary structure
mimicry by such species, protein-like tertiary folds remain elusive.
A fundamental challenge underlying this task is the design of a sequence
of side chains that will specify a complex tertiary folding pattern
on an unnatural backbone. We report here a sequence-based approach
to convert a natural protein with a compact tertiary fold to an analogue
with a backbone composed of ∼20% unnatural building blocks
but folding behavior similar to that of the parent protein
Introduction of Cyclically Constrained γ‑Residues Stabilizes an α‑Peptide Hairpin in Aqueous Solution
The synthesis and structural characterization of hybrid α/γ-peptides resulting from a 1:1 α→γ residue substitution at cross-strand positions in a hairpin-forming α-peptide sequence are described. Cyclically constrained γ-residues based on 1,3-substituted cyclohexane or benzene scaffolds support a native-like hairpin fold in aqueous solution, and the unnatural residues stabilize the folded state by ∼0.2 kcal/mol per α→γ substitution