Membrane interaction and secondary structure of de novo designed arginine-and tryptophan peptides with dual function

Rydberg, Hanna; Carlsson, Nils; Nordén, Bengt

doi:10.1016/j.bbrc.2012.09.030

Cited by 34 publications

(33 citation statements)

References 32 publications

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“…Globally, our findings are consistent with the results obtained by Rydberg et al [21,55], that is increasing the number of Trp leads to better uptake together with an increased cytotoxicity and membrane permeation, with a threshold at 4 Trp. Rydberg et al also show that the affinity of their peptides for membrane models all fall within the same range, making this parameter a bad predictor for cell uptake.…”

Section: Figuresupporting

confidence: 93%

“…On the one hand, Mitchell and Wender previously reported that decreasing the number of Arg from 9 to 5 in oligoarginine sequences led to a stepwise and linear decrease in internalization efficiency of the corresponding peptides [53,54]. On the other hand, Rydberg et al also investigated the number and position of Trp residues in polyarginine sequences, keeping the number of Arg residues constant (eight) and increasing the number of Trp stepwise [21,55]. Herein we kept the peptide length to 9 residues and replaced gradually Arg with Trp.…”

Section: Discussionmentioning

confidence: 99%

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Ionpair-π interactions favor cell penetration of arginine/tryptophan-rich cell-penetrating peptides

Walrant

Bauzá

Girardet

et al. 2019

Preprint

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Cell-penetrating peptides (CPPs) internalization can occur both by endocytosis and direct translocation through the cell membrane. These different entry routes suggest that molecular partners at the plasma membrane, phospholipids or glycosaminoglycans (GAGs), bind CPPs with different affinity or selectivity. The analysis of sequence-dependent interactions of CPPs with lipids and GAGs should lead to a better understanding of the molecular mechanisms underlying their internalization. CPPs are short sequences generally containing a high number of basic arginines and lysines and sometimes aromatic residues, in particular tryptophans. Tryptophans are crucial residues in membrane-active peptides, because they are important for membrane interaction. Membrane-active peptides often present facial amphiphilicity, which also promote the interaction with lipid bilayers. To study the role of Trp and facial amphiphilicity in cell interaction and penetration of CPPs, a nonapeptide series containing only Arg, Trp or D-Trp residues at different positions was designed. Our quantitative study indicates that to maintain/increase the uptake efficiency, Arg can be advantageously replaced by Trp in the nonapeptides. The presence of Trp in oligoarginines increases the uptake in cells expressing GAGs at their surface, when it only compensates for the loss of Arg and maintains similar peptide uptake in GAGdeficient cells. In addition, we show that facial amphiphilicity is not required for efficient uptake of these nonapeptides. Thermodynamic analyses point towards a key role of Trp that highly contributes to the binding enthalpy of complexes formation. Density functional theory (DFT) analysis highlights that salt bridge-π interactions play a crucial role for the GAG-dependent entry mechanisms. Peptide synthesisAll peptides were synthesized using standard Boc solid phase peptide synthesis. Boc-l-Arg(Tos), Boc-l-Trp(For), Boc-Gly, d-Biotin, MBHA Resin (0.53 mmol/g loading) and HBTU were purchased from Iris Biotech GmbH. Boc-(2,2-D 2 )-Gly was obtained from Cambridge Isotope Laboratories. Boc-d-Trp was purchased from Sigma-Aldrich. d-Biotin was fully oxidized to d-Biotin sulfone (Biot(O 2 ), Figure S1) by 4 days treatment with 30% H 2 O 2 in H 2 O and used without further purification. This avoids further oxidation of the peptide throughout time. Peptides were synthetized by hand on a 0.1 mmol scale for non-deuterated peptides and 0.01 mmol for deuterated peptides. Amino acid (5 eq) activation was performed by HBTU (4.5 eq) in the presence of excess DIEA (12 eq), and Boc deprotection was performed in neat TFA (2× 1 min). Trp side chains were deprotected prior to cleavage by treatment with 10% piperidine in DMF (1, 2, 5, 10, 30 and 60 min successive incubations). Peptides were cleaved from the resin by anhydrous HF (2h, 0°C) in the presence of anisole and dimethylsulfide.

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Section: Figuresupporting

confidence: 93%

Section: Discussionmentioning

confidence: 99%

Ionpair-π interactions favor cell penetration of arginine/tryptophan-rich cell-penetrating peptides

Walrant

Bauzá

Girardet

et al. 2019

Preprint

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“…In QCM-D, a mass deposition of peptide on the lipid surface is seen as a decrease in frequency, and changes in the viscoelastic properties of the lipid membrane upon peptide binding are related to changes in the dissipation signal. The peptide concentration of 5 µM, used for the surface interaction experiments, was chosen since the peptides at this concentration show evident biological responses in regard to both cellular uptake and bacterial growth inhibition, as seen in our previous results (Rydberg et al 2012a, b). …”

Section: Resultsmentioning

confidence: 99%

“…We have previously shown that the number and, even more interestingly, the position of tryptophan in the peptide sequence of a series of arginine-tryptophan peptides can affect both their uptake efficiency into live mammalian cells and their ability to inhibit bacterial growth (Rydberg et al 2012a, b). The uptake into cells was favored by an even distribution of tryptophans, while the peptides with an accumulation of tryptophans at the N-terminus of the peptide sequence seemed to be more efficient as antibacterial agents.…”

Section: Introductionmentioning

confidence: 99%

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Peptide-membrane interactions of arginine-tryptophan peptides probed using quartz crystal microbalance with dissipation monitoring

et al. 2014

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Membrane-active peptides include peptides that can cross cellular membranes and deliver macromolecular cargo as well as peptides that inhibit bacterial growth. Some of these peptides can act as both transporters and antibacterial agents. It is desirable to combine the knowledge from these two different fields of membrane-active peptides into design of new peptides with tailored actions, as transporters of cargo or as antibacterial substances, targeting specific membranes. We have previously shown that the position of the amino acid tryptophan in the peptide sequence of three arginine-tryptophan peptides affects their uptake and intracellular localization in live mammalian cells, as well as their ability to inhibit bacterial growth. Here, we use quartz crystal microbalance with dissipation monitoring to assess the induced changes caused by binding of the three peptides to supported model membranes composed of POPC, POPC/POPG, POPC/POPG/cholesterol or POPC/lactosyl PE. Our results indicate that the tryptophan position in the peptide sequence affects the way these peptides interact with the different model membranes and that the presence of cholesterol in particular seems to affect the membrane interaction of the peptide with an even distribution of tryptophans in the peptide sequence. These results give mechanistic insight into the function of these peptides and may aid in the design of membrane-active peptides with specified cellular targets and actions.Electronic supplementary materialThe online version of this article (doi:10.1007/s00249-014-0958-9) contains supplementary material, which is available to authorized users.

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Designing stapled peptides to inhibit protein‐protein interactions: An analysis of successes in a rapidly changing field

et al. 2020

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Two decades after their discovery, stapled peptide methodologies have evolved to a point where they can be used with confidence to generate therapeutic leads. Research groups across the world are testing innovative methodologies for their design, with dozens of publications released every month. A number of stapled peptide drug candidates have recently entered clinical trials. In this review, we provide an overview of successful methods for their construction, highlight trends in the deposited crystal structures of stapled peptide complexed to their targets and discuss properties that contribute towards improved pharmacological profiles.

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Membrane interaction and secondary structure of de novo designed arginine-and tryptophan peptides with dual function

Cited by 34 publications

References 32 publications

Ionpair-π interactions favor cell penetration of arginine/tryptophan-rich cell-penetrating peptides

Ionpair-π interactions favor cell penetration of arginine/tryptophan-rich cell-penetrating peptides

Peptide-membrane interactions of arginine-tryptophan peptides probed using quartz crystal microbalance with dissipation monitoring

Designing stapled peptides to inhibit protein‐protein interactions: An analysis of successes in a rapidly changing field

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