EJP18 peptide derived from the juxtamembrane domain of epidermal growth factor receptor represents a novel membrane-active cell-penetrating peptide

N.G. Eissa; E.J. Sayers; D. Birch; S.G. Patel; Y.-H. Tsai; H. Mørck Nielsen; A.T. Jones

doi:10.1042/bcj20190452

EJP18 peptide derived from the juxtamembrane domain of epidermal growth factor receptor represents a novel membrane-active cell-penetrating peptide

Biochemical Journal ◽

10.1042/bcj20190452 ◽

2020 ◽

Vol 477 (1) ◽

pp. 45-60 ◽

Cited By ~ 1

Author(s):

N.G. Eissa ◽

E.J. Sayers ◽

D. Birch ◽

S.G. Patel ◽

Y.-H. Tsai ◽

...

Keyword(s):

Antimicrobial Agents ◽

Cell Penetrating Peptides ◽

Cell Physiology ◽

Cellular Delivery ◽

Membrane Interactions ◽

Active Peptides ◽

Juxtamembrane Region ◽

Protein Membrane ◽

Cell Penetrating ◽

Membrane Active Peptides

Membrane-active peptides have been extensively studied to probe protein–membrane interactions, to act as antimicrobial agents and cell-penetrating peptides (CPPs) for the delivery of therapeutic agents to cells. Hundreds of membrane-active sequences acting as CPPs have now been described including bioportides that serve as single entity modifiers of cell physiology at the intracellular level. Translation of promising CPPs in pre-clinical studies have, however, been disappointing as only few identified delivery systems have progressed to clinical trials. To search for novel membrane-active peptides a sequence from the EGFR juxtamembrane region was identified (named EJP18), synthesised, and examined in its L- and D-form for its ability to mediate the delivery of a small fluorophore and whole proteins to cancer cell lines. Initial studies identified the peptide as being highly membrane-active causing extensive and rapid plasma membrane reorganisation, blebbing, and toxicity. At lower, non-toxic concentrations the peptides outperformed the well-characterised CPP octaarginine in cellular delivery capacity for a fluorophore or proteins that were associated with the peptide covalently or via ionic interactions. EJP18 thus represents a novel membrane-active peptide that may be used as a naturally derived model for biophysical protein–membrane interactions or for delivery of cargo into cells for therapeutic or diagnostic applications.

Download Full-text

Membrane-Active Peptides: Stable Pore-Forming or Cell-Penetrating Peptides Selected With Orthogonal High-Throughput Screening

Biophysical Journal ◽

10.1016/j.bpj.2009.12.483 ◽

2010 ◽

Vol 98 (3) ◽

pp. 85a

Author(s):

Jessica R. Marks ◽

Aram J. Krauson ◽

Kalina A. Hristova ◽

William C. Wimley

Keyword(s):

High Throughput ◽

High Throughput Screening ◽

Cell Penetrating Peptides ◽

Active Peptides ◽

Cell Penetrating ◽

Membrane Active Peptides

Download Full-text

Membrane Active Peptides and Their Biophysical Characterization

Biomolecules ◽

10.3390/biom8030077 ◽

2018 ◽

Vol 8 (3) ◽

pp. 77 ◽

Cited By ~ 41

Author(s):

Fatma Gizem Avci ◽

Berna Sariyar Akbulut ◽

Elif Ozkirimli

Keyword(s):

Antimicrobial Peptides ◽

Single Molecule ◽

Imaging Techniques ◽

Membrane Integrity ◽

Cell Penetrating Peptides ◽

Single Molecule Level ◽

Active Peptides ◽

Cell Penetrating ◽

Biophysical Techniques ◽

Membrane Active Peptides

In the last 20 years, an increasing number of studies have been reported on membrane active peptides. These peptides exert their biological activity by interacting with the cell membrane, either to disrupt it and lead to cell lysis or to translocate through it to deliver cargos into the cell and reach their target. Membrane active peptides are attractive alternatives to currently used pharmaceuticals and the number of antimicrobial peptides (AMPs) and peptides designed for drug and gene delivery in the drug pipeline is increasing. Here, we focus on two most prominent classes of membrane active peptides; AMPs and cell-penetrating peptides (CPPs). Antimicrobial peptides are a group of membrane active peptides that disrupt the membrane integrity or inhibit the cellular functions of bacteria, virus, and fungi. Cell penetrating peptides are another group of membrane active peptides that mainly function as cargo-carriers even though they may also show antimicrobial activity. Biophysical techniques shed light on peptide–membrane interactions at higher resolution due to the advances in optics, image processing, and computational resources. Structural investigation of membrane active peptides in the presence of the membrane provides important clues on the effect of the membrane environment on peptide conformations. Live imaging techniques allow examination of peptide action at a single cell or single molecule level. In addition to these experimental biophysical techniques, molecular dynamics simulations provide clues on the peptide–lipid interactions and dynamics of the cell entry process at atomic detail. In this review, we summarize the recent advances in experimental and computational investigation of membrane active peptides with particular emphasis on two amphipathic membrane active peptides, the AMP melittin and the CPP pVEC.

Download Full-text

Membrane-Active Peptides from Marine Organisms—Antimicrobials, Cell-Penetrating Peptides and Peptide Toxins: Applications and Prospects

Probiotics and Antimicrobial Proteins ◽

10.1007/s12602-014-9182-2 ◽

2015 ◽

Vol 7 (1) ◽

pp. 75-89 ◽

Cited By ~ 18

Author(s):

Nisha Ponnappan ◽

Deepthi Poornima Budagavi ◽

Bhoopesh Kumar Yadav ◽

Archana Chugh

Keyword(s):

Cell Penetrating Peptides ◽

Marine Organisms ◽

Active Peptides ◽

Cell Penetrating ◽

Peptide Toxins ◽

Membrane Active Peptides

Download Full-text

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

10.1101/717207 ◽

2019 ◽

Cited By ~ 1

Author(s):

Astrid Walrant ◽

Antonio Bauzá ◽

Claudia Girardet ◽

Isabel D. Alves ◽

Sophie Lecomte ◽

...

Keyword(s):

Lipid Bilayers ◽

Density Functional ◽

Molecular Mechanisms ◽

Cell Penetrating Peptides ◽

Membrane Phospholipids ◽

Active Peptides ◽

Π Interactions ◽

Cell Penetrating ◽

Membrane Active Peptides

AbstractCell-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 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.

Download Full-text

Cellular Delivery of Peptide Nucleic Acid by Cell-Penetrating Peptides

Peptide Synthesis and Applications ◽

10.1385/1-59259-877-3:131 ◽

2005 ◽

pp. 131-141 ◽

Cited By ~ 1

Author(s):

Kilk Kalle ◽

lo Langel

Keyword(s):

Nucleic Acid ◽

Peptide Nucleic Acid ◽

Cell Penetrating Peptides ◽

Cellular Delivery ◽

Cell Penetrating

Download Full-text

In Vitro Activities of Membrane-Active Peptides Alone and in Combination with Clinically Used Antimicrobial Agents against Stenotrophomonas maltophilia

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.44.6.1716-1719.2000 ◽

2000 ◽

Vol 44 (6) ◽

pp. 1716-1719 ◽

Cited By ~ 28

Author(s):

A. Giacometti ◽

O. Cirioni ◽

M. S. Del Prete ◽

F. Barchiesi ◽

M. Fortuna ◽

...

Keyword(s):

Stenotrophomonas Maltophilia ◽

Antimicrobial Agents ◽

Antimicrobial Activities ◽

Active Peptides ◽

Cecropin P1 ◽

Polymyxin E ◽

Membrane Active Peptides ◽

Magainin Ii ◽

Time Kill

ABSTRACT The in vitro activities of buforin II, cecropin P1, and magainin II, alone and in combination with six clinically used antimicrobial agents, against 12 clinical isolates of Stenotrophomonas maltophilia were investigated. Antimicrobial activities were measured by MIC and time-kill studies. The isolates were susceptible to the peptides at concentrations in the range of 0.50 to 16 μg/ml. Synergy was observed when the peptides were combined with polymyxin E, meropenem, ceftazidime, piperacillin, and clarithromycin.

Download Full-text

Cell‐penetrating peptides recruit type A scavenger receptors to the plasma membrane for cellular delivery of nucleic acids

The FASEB Journal ◽

10.1096/fj.201600811r ◽

2016 ◽

Vol 31 (3) ◽

pp. 975-988 ◽

Cited By ~ 11

Author(s):

Carmen Juks ◽

Annely Lorents ◽

Piret Arukuusk ◽

Ülo Langel ◽

Margus Pooga

Keyword(s):

Plasma Membrane ◽

Nucleic Acids ◽

Type A ◽

Cell Penetrating Peptides ◽

Scavenger Receptors ◽

Cellular Delivery ◽

Cell Penetrating

Download Full-text

Membrane Active Peptides and Their Biophysical Characterization

10.20944/preprints201807.0008.v1 ◽

2018 ◽

Author(s):

Fatma Gizem Avci ◽

Berna Sariyar Akbulut ◽

Elif Ozkirimli

Keyword(s):

Single Molecule ◽

Imaging Techniques ◽

Membrane Integrity ◽

Drug Market ◽

Cell Penetrating Peptides ◽

Single Molecule Level ◽

Active Peptides ◽

Biophysical Techniques ◽

Membrane Active Peptides ◽

Dynamics Simulations

In the last 20 years, an increasing number of studies have been reported on membrane active peptides, which exert their biological activity by interacting with the cell membrane either to disrupt it and lead to cell lysis or to translocate through it to deliver cargos into the cell and reach their target. These peptides are attractive alternatives to currently used pharmaceuticals. Antimicrobial peptides (AMPs) and peptides designed for drug and gene delivery currently in the drug pipeline suggest that these membrane active peptides will soon constitute a significant percentage of the drug market. Here, we focus on two most prominent classes of membrane active peptides; AMPs and cell-penetrating peptides (CPPs). AMPs are a group of membrane active peptides that disrupt the membrane integrity or inhibit the cellular functions of bacteria, virus and fungi. CPPs are another group of membrane active peptides that mainly function as cargo-carriers even though they may also show antimicrobial activity to some extent. Biophysical techniques to understand how they interact with the membrane have shed light on the peptide–membrane interaction at various levels of detail. Structural investigation of membrane active peptides in the presence of the membrane provides important clues on the effect of the membrane environment on peptide conformations. Advances in live imaging techniques have allowed examination of peptide action at a single cell or single molecule level. In addition to these experimental biophysical techniques, molecular dynamics simulations provided clues on the peptide-lipid interactions and dynamics of the cell entry process at atomic detail. In this review, we summarize the recent advances in experimental and computational investigation of membrane active peptides with particular emphasis on two amphipathic membrane active peptides, the AMP melittin and the CPP pVEC.

Download Full-text