Membrane anchoring of a curvature-inducing peptide, EpN18, promotes membrane translocation of octaarginine

Kuroki, Kakeru; Sakai, Takayuki; Masuda, Takeo; Kawano, Kenichi; Futaki, Shiroh

doi:10.1016/j.bmcl.2021.128103

Cited by 7 publications

(6 citation statements)

References 14 publications

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“…In this inkjet system, the controlled jet of droplets with CPPs on targeted cells successfully showed membrane penetration and cytosolic delivery of macromolecules without disruption of plasma membranes. The formation of positive membrane curvature is crucial for membrane penetration of arginine-rich CPPs, and treatment of epsin-1 N-terminal peptide (EpN18), which can increase positive membrane curvature, promotes direct penetration of the CPPs through plasma membranes. − Droplets with the CPPs jetted using the inkjet system might induce membrane curvatures on plasma membranes because of the pressure of jetted droplets without any pore formations, thereby enhancing their membrane penetration and cytosolic release (Figure ). However, detailed mechanisms should be studied for understanding the system advantages from the viewpoints of biophysics.…”

Section: Discussionmentioning

confidence: 99%

Inkjet-Based Intracellular Delivery System that Effectively Utilizes Cell-Penetrating Peptides for Cytosolic Introduction of Biomacromolecules through the Cell Membrane

Omura,

Morimoto,

Araki

et al. 2023

ACS Appl. Mater. Interfaces

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In the drug delivery system, the cytosolic delivery of biofunctional molecules such as enzymes and genes must achieve sophisticated activities in cells, and microinjection and electroporation systems are typically used as experimental techniques. These methods are highly reliable, and they have high intracellular transduction efficacy. However, a high degree of proficiency is necessary, and induced cytotoxicity is considered as a technical problem. In this research, a new intracellular introduction technology was developed through the cell membrane using an inkjet device and cell-penetrating peptides (CPPs). Using the inkjet system, the droplet volume, droplet velocity, and dropping position can be accurately controlled, and minute samples (up to 30 pL/shot) can be carried out by direct administration. In addition, CPPs, which have excellent cell membrane penetration functions, can deliver high-molecular-weight drugs and nanoparticles that are difficult to penetrate through the cell membrane. By using the inkjet system, the CPPs with biofunctional cargo, including peptides, proteins such as antibodies, and exosomes, could be accurately delivered to cells, and efficient cytosolic transduction was confirmed.

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Section: Discussionmentioning

confidence: 99%

Inkjet-Based Intracellular Delivery System that Effectively Utilizes Cell-Penetrating Peptides for Cytosolic Introduction of Biomacromolecules through the Cell Membrane

Omura,

Morimoto,

Araki

et al. 2023

ACS Appl. Mater. Interfaces

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“…Original EpN18 has a predicted α-helical structure (Figure 1A: XSTSSLRRQXKNIVHNYS-amide, where X = norleucine/Nle substitutes native methionine to avoid ambiguity in polarity upon sulfur oxidation). [10] EpN18 contains six hydrophobic amino acids, Nle 1 , Leu 6 , Nle 10 , Ile 13 , Val 14 , and Tyr 17 (Figure 1A). As seen in the helical wheel projection, the latter five residues constitute the hydrophobic face of an amphipathic helix.…”

Section: Design Of Simplified Peptidesmentioning

confidence: 99%

“… [11a] By elucidating the underlying structural basis of how EpN18 is able to modulate membrane curvature, new peptide‐based tools can be generated for intentionally controlling numerous cellular phenomena without any change in lipid composition. Along this line, we have recently shown that multimerization and membrane anchoring of EpN18 leads to an enhanced interaction with membranes [12,13] . However, the structural features that specifically contribute to curvature induction and membrane sculpturing by EpN18 remain elusive.…”

Section: Introductionmentioning

confidence: 99%

“…Along this line, we have recently shown that multimerization and membrane anchoring of EpN18 leads to an enhanced interaction with membranes. [12,13] However, the structural features that specifically contribute to curvature induction and membrane sculpturing by EpN18 remain elusive. It has been reported that various cellular proteins involved in regulating membrane curvature likely share a common curvature-inducing mechanism, namely through the interaction of N-terminal amphiphilic helices with the membranes.…”

Section: Introductionmentioning

confidence: 99%

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Structural Dissection of Epsin‐1 N‐Terminal Helical Peptide: The Role of Hydrophobic Residues in Modulating Membrane Curvature

Nishimura

Kawaguchi

Kuroki

et al. 2023

Chemistry A European J

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Spatiotemporal structural alterations in cellular membranes are the hallmark of many vital processes. In these cellular events, the induction of local changes in membrane curvature often plays a pivotal role. Many amphiphilic peptides are able to modulate membrane curvature, but there is little information on specific structural factors that direct the curvature change. Epsin‐1 is a representative protein thought to initiate invagination of the plasma membrane upon clathrin‐coated vesicles formation. Its N‐terminal helical segment (EpN18) plays a key role in inducing positive membrane curvature. This study aimed to elucidate the essential structural features of EpN18 in order to better understand general curvature‐inducing mechanisms, and to design effective tools for rationally controlling membrane curvature. Structural dissection of peptides derived from EpN18 revealed the decisive contribution of hydrophobic residues to (i) enhancing membrane interactions, (ii) helix structuring, (iii) inducing positive membrane curvature, and (iv) loosening lipid packing. The strongest effect was obtained by substitution with leucine residues, as this EpN18 analog showed a marked ability to promote the influx of octa‐arginine cell‐penetrating peptides into living cells.

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“…Furthermore, we synthesized 40A-8R, a peptide composed of 40 amino acids of the effector site of MTI-II fused with 8 arginine residues in the C-terminus ( 36 ). Oligoarginines were added to amino acids because arginine-rich peptides can be efficiently internalized into cells, and they are widely used as carriers for the intracellular delivery of bioactive molecules ( 37 – 39 ).…”

Section: Introductionmentioning

confidence: 99%

Anti–NF-κB peptide derived from nuclear acidic protein attenuates ovariectomy-induced osteoporosis in mice

Takami,

Okamoto,

Etani

et al. 2023

JCI Insight

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NF-κB is a transcription factor that is activated with aging. It plays a key role in the development of osteoporosis by promoting osteoclast differentiation and inhibiting osteoblast differentiation. In this study, we developed a small anti–NF-κB peptide called 6A-8R from a nuclear acidic protein (also known as macromolecular translocation inhibitor II, Zn 2+ -binding protein, or parathymosin) that inhibits transcriptional activity of NF-κB without altering its nuclear translocation and binding to DNA. Intraperitoneal injection of 6A-8R attenuated ovariectomy-induced osteoporosis in mice by inhibiting osteoclast differentiation, promoting osteoblast differentiation, and inhibiting sclerostin production by osteocytes in vivo with no apparent side effects. Conversely, in vitro, 6A-8R inhibited osteoclast differentiation by inhibiting NF-κB transcriptional activity, promoted osteoblast differentiation by promoting Smad1 phosphorylation, and inhibited sclerostin expression in osteocytes by inhibiting myocyte enhancer factors 2C and 2D. These findings suggest that 6A-8R has the potential to be an antiosteoporotic therapeutic agent with uncoupling properties.

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Membrane anchoring of a curvature-inducing peptide, EpN18, promotes membrane translocation of octaarginine

Cited by 7 publications

References 14 publications

Inkjet-Based Intracellular Delivery System that Effectively Utilizes Cell-Penetrating Peptides for Cytosolic Introduction of Biomacromolecules through the Cell Membrane

Inkjet-Based Intracellular Delivery System that Effectively Utilizes Cell-Penetrating Peptides for Cytosolic Introduction of Biomacromolecules through the Cell Membrane

Structural Dissection of Epsin‐1 N‐Terminal Helical Peptide: The Role of Hydrophobic Residues in Modulating Membrane Curvature

Anti–NF-κB peptide derived from nuclear acidic protein attenuates ovariectomy-induced osteoporosis in mice

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