Photoswitching of Cell Penetration of Amphipathic Peptides by Control of α-Helical Conformation

Kim, Gyu Chan; Ahn, Joon Hyung; Oh, Jin-Seok; Nam, Sohee; Hyun, Soonsil; Yu, Jaehoon; Lee, Yan

doi:10.1021/acs.biomac.8b00428

Cited by 32 publications

(30 citation statements)

References 49 publications

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“…However, it later appeared that amphipathicity had no direct impact on internalization of the KLA peptide, but could promote binding to intracellular organelle membranes. Strategies exploiting switchable and/or structurally constrained amphipathic helical peptides to control or enhance uptake are still being developed [26][27][28]. CPPs are often classified according to their ability to adopt a secondary amphipathic structure [23,29,30] possibly by analogy with cationic α-helical antimicrobial peptides, another class of membrane-active peptides [31,32].…”

Section: Introductionmentioning

confidence: 99%

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

Walrant

Bauzá

Girardet

et al. 2020

Biochimica et Biophysica Acta (BBA) - Biomembranes

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Cell-penetrating peptides (CPPs) internalization 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, while it compensates for the loss of charge interactions from Arg and maintains similar peptide uptake in GAG-deficient 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.

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Section: Introductionmentioning

confidence: 99%

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

Walrant

Bauzá

Girardet

et al. 2020

Biochimica et Biophysica Acta (BBA) - Biomembranes

View full text Add to dashboard Cite

show abstract

Section: Introductionmentioning

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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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: Main Specific Applications Of Azobenzenementioning

confidence: 99%

“…The use of azobenzene to control the formation of α‐helical conformations to regulate permeabilization is a good example, which proves the applicability of this molecular switch in biomedical applications. Lee and co‐workers [ 234 ] used an azobenzene linker to modulate the secondary structure of peptides (Figure 14e). The azobenzene linker is connected with an amphipathic peptide which is chiefly consisting of leucine (L) and lysine (K) (LK peptide, a cell‐penetrating peptides).…”

Section: Main Specific Applications Of Azobenzenementioning

confidence: 99%

Advances in Application of Azobenzene as a Trigger in Biomedicine: Molecular Design and Spontaneous Assembly

et al. 2021

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Azobenzene is a well‐known derivative of stimulus‐responsive molecular switches and has shown superior performance as a functional material in biomedical applications. The results of multiple studies have led to the development of light/hypoxia‐responsive azobenzene for biomedical use. In recent years, long‐wavelength‐responsive azobenzene has been developed. Matching the longer wavelength absorption and hypoxia‐response characteristics of the azobenzene switch unit to the bio‐optical window results in a large and effective stimulus response. In addition, azobenzene has been used as a hypoxia‐sensitive connector via biological cleavage under appropriate stimulus conditions. This has resulted in on/off state switching of properties such as pharmacology and fluorescence activity. Herein, recent advances in the design and fabrication of azobenzene as a trigger in biomedicine are summarized.

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Photoswitching of Cell Penetration of Amphipathic Peptides by Control of α-Helical Conformation

Cited by 32 publications

References 49 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

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

Advances in Application of Azobenzene as a Trigger in Biomedicine: Molecular Design and Spontaneous Assembly

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