Single Turn Peptide Alpha Helices with Exceptional Stability in Water

Shepherd, Nicholas E.; Hoang, Huy N.; Abbenante, Giovanni; Fairlie, David P.

doi:10.1021/ja0456003

Cited by 304 publications

(315 citation statements)

References 93 publications

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“…In agreement with our earlier study of small model peptides (11), CD spectral fingerprints show that the α-helical pentapeptide (1) is stable to aqueous phosphate buffer, acid, base, trypsin degradation, does not unfold in a denaturing environment (e.g., 8 M guanidine), and the CD spectral molar ellipticity does not increase at 222 nm in 50% 2,2,2-trifluoroethanol (TFE) indicating that maximum helicity is enforced by the constraint (Fig. 1B and Table S5).…”

Section: Resultssupporting

confidence: 93%

“…The 116 residue HIV-Rev protein binds to RNA with Kd approximately 1 nM (28). Nociceptin (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17) is an agonist at only nM concentrations (25). We also found the helix-constrained compounds to be more stable in human serum than their linear analogues, the latter being typically degraded within 1 h whereas the constrained peptides were stable for >24 h, except for the exocyclic residues, which were cleaved by proteases over several hours.…”

Section: Discussionmentioning

confidence: 90%

“…We previously demonstrated that arginine-containing pentapeptides Ac-(1,5-cyclo)-[KXRXD]-NH 2 , and related hexapeptides featuring an amide bond linkage between lysine and aspartate side chains spaced three residues apart, were α-helical by CD spectroscopy (11). However, CD spectra are notoriously unreliable for characterising α-helicity in short peptides (11)(12)(13). Here we report the NMR solution structure of Ac-(1,5-cyclo)-[KAAAD]-NH 2 (1), containing three alanines, that is unambiguously the smallest three dimensional α-helical structure ever reported in water.…”

Section: Resultsmentioning

confidence: 99%

“…This relatively low helicity for mimetic 26 versus mimetics 20, 22, and 24 is attributed to the unstructured "triggering" message domain at the N-terminus, as well as accumulation of positive charges on one helix face. Nevertheless, the induction of at least some helix structure in this peptide suggested that it should be more active than nociceptin (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17). Function was investigated by nociceptin(1-17) induced intracellular ERK phosphorylation in mouse Neuro-2a neuroblastoma cells (Fig.…”

Section: Resultsmentioning

confidence: 99%

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Downsizing human, bacterial, and viral proteins to short water-stable alpha helices that maintain biological potency

Harrison

Shepherd²,

Hoang³

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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171

166

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Recombinant proteins are important therapeutics due to potent, highly specific, and nontoxic actions in vivo. However, they are expensive medicines to manufacture, chemically unstable, and difficult to administer with low patient uptake and compliance. Small molecule drugs are cheaper and more bioavailable, but less target-specific in vivo and often have associated side effects. Here we combine some advantages of proteins and small molecules by taking short amino acid sequences that confer potency and selectivity to proteins, and fixing them as small constrained molecules that are chemically and structurally stable and easy to make. Proteins often use short α-helices of just 1-4 helical turns (4-15 amino acids) to interact with biological targets, but peptides this short usually have negligible α-helicity in water. Here we show that short peptides, corresponding to helical epitopes from viral, bacterial, or human proteins, can be strategically fixed in highly α-helical structures in water. These helix-constrained compounds have similar biological potencies as proteins that bear the same helical sequences. Examples are (i) a picomolar inhibitor of Respiratory Syncytial Virus F protein mediated fusion with host cells, (ii) a nanomolar inhibitor of RNA binding to the transporter protein HIV-Rev, (iii) a submicromolar inhibitor of Streptococcus pneumoniae growth induced by quorum sensing pheromone Competence Stimulating Peptide, and (iv) a picomolar agonist of the GPCR pain receptor opioid receptor like receptor ORL-1. This approach can be generally applicable to downsizing helical regions of proteins with broad applications to biology and medicine.helix | inhibitor | structure | mimetic | anti-infective P roteins are key functional components that define life, aging, disease, and death. Their uses in medicine, science, and industry are however limited by their complexity, high costs, chemical instability, and low bioavailability. Consequently, new chemical technology is needed to create simpler, smaller, cheaper, more stable, and bioavailable molecules that can execute or inhibit selected functions of proteins. If generic approaches could be developed to stabilize or recreate new molecular shapes that mimic structural components of proteins (e.g., helices, turns, strands, and combinations), the resulting molecules could potentially be extremely valuable for interrogating biological systems and for exploring prospective applications such as new pharmaceuticals, diagnostics, vaccines, and nanomaterials.Some proteins elicit biological function through a single short α-helical segment (usually 4-15 amino acids in 1-4 helical turns) that interacts with nucleic acids or proteins (1-3). However, short synthetic peptides of this length are usually not thermodynamically stable helices in water and adopt only random structures (4). Techniques developed as generalized strategies for stabilising or mimicking short peptide α-helices have been limited by lack of helical structure in water, by sequence or context dependent helici...

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Section: Resultssupporting

confidence: 93%

Section: Discussionmentioning

confidence: 90%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

See 2 more Smart Citations

Downsizing human, bacterial, and viral proteins to short water-stable alpha helices that maintain biological potency

Harrison

Shepherd²,

Hoang³

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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171

166

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show abstract

“…Otherwise, efficient macrocyclization of linear peptides through the formation of an oxadiazole has also been reported 8. However, a large number of stapling/macrocyclization/re‐bridging strategies consist of the introduction of non‐canonical amino acids and their subsequent ligation by ring‐closing metathesis,9 lactamization,10 or cycloaddition reactions 2c, 4b. Common to many of these strategies is either the requirement for complicated orthogonal protection procedures, sequence engineering, the appendage of bulky/constrained linkers between the two residues, or the use of organic solvents.…”

mentioning

confidence: 99%

Oxetane Grafts Installed Site‐Selectively on Native Disulfides to Enhance Protein Stability and Activity In Vivo

et al. 2017

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A four‐membered oxygen ring (oxetane) can be readily grafted into native peptides and proteins through site‐selective bis‐alkylation of cysteine residues present as disulfides under mild and biocompatible conditions. The selective installation of the oxetane graft enhances stability and activity, as demonstrated for a range of biologically relevant cyclic peptides, including somatostatin, proteins, and antibodies, such as a Fab arm of the antibody Herceptin and a designed antibody DesAb‐Aβ against the human Amyloid‐β peptide. Oxetane grafting of the genetically detoxified diphtheria toxin CRM197 improves significantly the immunogenicity of this protein in mice, which illustrates the general utility of this strategy to modulate the stability and biological activity of therapeutic proteins containing disulfides in their structures.

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Peptidomimetics of α‐Helical and β‐Strand Protein Binding Epitopes

Bionda

Fasan

2015

Small Molecule Medicinal Chemistry

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Single Turn Peptide Alpha Helices with Exceptional Stability in Water

Cited by 304 publications

References 93 publications

Downsizing human, bacterial, and viral proteins to short water-stable alpha helices that maintain biological potency

Downsizing human, bacterial, and viral proteins to short water-stable alpha helices that maintain biological potency

Oxetane Grafts Installed Site‐Selectively on Native Disulfides to Enhance Protein Stability and Activity In Vivo

Peptidomimetics of α‐Helical and β‐Strand Protein Binding Epitopes

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