Lysine to arginine mutagenesis of chlorotoxin enhances its cellular uptake

Ojeda, Paola G.; Henriques, Sónia Troeira; Pan, Yijun; Nicolazzo, Joseph A.; Craik, David J.; Wang, Conan K.

doi:10.1002/bip.23025

Cited by 13 publications

(10 citation statements)

References 69 publications

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“…The latter peptide had stronger activity than the all-lysine one against both promastigotes and amastigotes of the different species/strains of Leishmania parasites tested (27 to 71 µM for promastigotes; 16 to 18 µM for amastigotes); moreover, p–Ac1R7 displayed a faster parasite membrane-permeation action than p–Ac1 [ 42 ]. These observations are in line with studies involving antimicrobial and anticancer peptides that have shown a greater capacity to penetrate cells when having a high density of arginine residues, as compared to analogs of the same length containing lysines instead [ 85 , 86 ].…”

Section: Understanding the Primary Structure And Composition Of Leishmanicidal Peptidessupporting

confidence: 82%

Peptides to Tackle Leishmaniasis: Current Status and Future Directions

Robles-Loaiza

Pinos-Tamayo

Mendes

et al. 2021

IJMS

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Peptide-based drugs are an attractive class of therapeutic agents, recently recognized by the pharmaceutical industry. These molecules are currently being used in the development of innovative therapies for diverse health conditions, including tropical diseases such as leishmaniasis. Despite its socioeconomic influence on public health, leishmaniasis remains long-neglected and categorized as a poverty-related disease, with limited treatment options. Peptides with antileishmanial effects encountered to date are a structurally heterogeneous group, which can be found in different natural sources—amphibians, reptiles, insects, bacteria, marine organisms, mammals, plants, and others—or inspired by natural toxins or proteins. This review details the biochemical and structural characteristics of over one hundred peptides and their potential use as molecular frameworks for the design of antileishmanial drug leads. Additionally, we detail the main chemical modifications or substitutions of amino acid residues carried out in the peptide sequence, and their implications in the development of antileishmanial candidates for clinical trials. Our bibliographic research highlights that the action of leishmanicidal peptides has been evaluated mainly using in vitro assays, with a special emphasis on the promastigote stage. In light of these findings, and considering the advances in the successful application of peptides in leishmaniasis chemotherapy, possible approaches and future directions are discussed here.

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Section: Understanding the Primary Structure And Composition Of Leishmanicidal Peptidessupporting

confidence: 82%

Peptides to Tackle Leishmaniasis: Current Status and Future Directions

Robles-Loaiza

Pinos-Tamayo

Mendes

et al. 2021

IJMS

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“…Accordingly, cellular uptake of TM601 in U373 glioma cells was a rapid concentration-and time-dependent process, which was affected by chlorpromazine, a pharmacological inhibitor of clathrin-mediated endocytosis, involved in the transport of coated pits. Interestingly, as in the case of lycosin-I from spider venom, the replacement of Lys with Arg in the CTX sequence, as observed with the synthetic analogs [K15R/K23R]CTX and [K15R/K23R/Y29W], enhanced its cellular uptake, as demonstrated in human cervical carcinoma (HeLa) cells with fluorescent derivatives [95].…”

Section: Chlorotoxinmentioning

confidence: 81%

Cell-Penetrating Peptides Derived from Animal Venoms and Toxins

Rádis‐Baptista

2021

Toxins

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Cell-penetrating peptides (CPPs) comprise a class of short polypeptides that possess the ability to selectively interact with the cytoplasmic membrane of certain cell types, translocate across plasma membranes and accumulate in the cell cytoplasm, organelles (e.g., the nucleus and mitochondria) and other subcellular compartments. CPPs are either of natural origin or de novo designed and synthesized from segments and patches of larger proteins or designed by algorithms. With such intrinsic properties, along with membrane permeation, translocation and cellular uptake properties, CPPs can intracellularly convey diverse substances and nanomaterials, such as hydrophilic organic compounds and drugs, macromolecules (nucleic acids and proteins), nanoparticles (nanocrystals and polyplexes), metals and radionuclides, which can be covalently attached via CPP N- and C-terminals or through preparation of CPP complexes. A cumulative number of studies on animal toxins, primarily isolated from the venom of arthropods and snakes, have revealed the cell-penetrating activities of venom peptides and toxins, which can be harnessed for application in biomedicine and pharmaceutical biotechnology. In this review, I aimed to collate examples of peptides from animal venoms and toxic secretions that possess the ability to penetrate diverse types of cells. These venom CPPs have been chemically or structurally modified to enhance cell selectivity, bioavailability and a range of target applications. Herein, examples are listed and discussed, including cysteine-stabilized and linear, α-helical peptides, with cationic and amphipathic character, from the venom of insects (e.g., melittin, anoplin, mastoparans), arachnids (latarcin, lycosin, chlorotoxin, maurocalcine/imperatoxin homologs and wasabi receptor toxin), fish (pardaxins), amphibian (bombesin) and snakes (crotamine and cathelicidins).

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“…However, given that CTX-3 binds to both a tumor cell line and a normal cell line, in contrast to previous studies on CTX, it appears likely that this region is not sufficient by itself to account for the selective binding to tumor cells. It is possible the cell surface binding observed for CTX-3 is non-specific membrane binding mediated by the presence of the high proportion of positively charged (two lysine and one arginine) residues within a seven-residue peptide as reported for other highly charged peptides (Futaki et al., 2007; Ojeda et al., 2017). Although no cell surface binding was observed for CTX-4, it was the only fragment that appeared to be specifically internalized into the glioma cells at 37°C.…”

Section: Discussionmentioning

confidence: 92%

A C-Terminal Fragment of Chlorotoxin Retains Bioactivity and Inhibits Cell Migration

et al. 2019

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Chlorotoxin was originally isolated from the venom of the Israeli scorpion Leiurus quinquestriatus, and has potential as a tumor imaging agent based on its selective binding to tumor cells. Several targets have been suggested for chlorotoxin including voltage-gated chloride channels, and it has been shown to have anti-angiogenic activity and inhibit cell migration. The structure of chlorotoxin is stabilized by four disulfide bonds and contains β-sheet and helical structure. Interestingly, the reduced form has previously been shown to inhibit cell migration to the same extent as the wild type, but structural analysis indicates that the reduced form of the peptide does not maintain the native secondary structure and appears unstructured in solution. This lack of structure suggests that a short stretch of amino acids might be responsible for the bioactivity. To explore this hypothesis, we have synthesized fragments of chlorotoxin without disulfide bonds. As expected for such small peptides, NMR analysis indicated that the peptides were unstructured in solution. However, the peptide corresponding to the eight C-terminal residues inhibited cell migration, in contrast to the other fragments. Our results suggest that the C-terminal region plays a critical role in the bioactivity of chlorotoxin.

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Lysine to arginine mutagenesis of chlorotoxin enhances its cellular uptake

Cited by 13 publications

References 69 publications

Peptides to Tackle Leishmaniasis: Current Status and Future Directions

Peptides to Tackle Leishmaniasis: Current Status and Future Directions

Cell-Penetrating Peptides Derived from Animal Venoms and Toxins

A C-Terminal Fragment of Chlorotoxin Retains Bioactivity and Inhibits Cell Migration

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