Automated NMR structure determination and disulfide bond identification of the myotoxin crotamine from Crotalus durissus terrificus

Fadel, Valmir; Bettendorff, Pascal; Herrmann, Torsten; Azevedo, Walter Filgueira de; Oliveira, Eduardo B.; Yamane, Toshimi; Wüthrich, Kurt

doi:10.1016/j.toxicon.2005.07.018

Cited by 82 publications

(59 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The band in the region 221-231 is consistent with a B b transition arising from Trp residues and/or with L a (031) transition associated with the Tyr residues. Alternatively, this band can arise from the existing disulfide bonds in the crotamine structure between residues Cys-4 -Cys-36, Cys-11-Cys-30, and Cys-18 -Cys-37 (19,20). The observed spectral changes caused by the presence of heparin are indicative of significant alteration in the secondary structure of crotamine.…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Crotamine Mediates Gene Delivery into Cells through the Binding to Heparan Sulfate Proteoglycans

Nascimento

Hayashi

Kerkis

et al. 2007

Journal of Biological Chemistry

Self Cite

104

119

View full text Add to dashboard Cite

Recently we have shown that crotamine, a toxin from the South American rattlesnake Crotalus durissus terrificus venom, belongs to the family of cell-penetrating peptides. Moreover, crotamine was demonstrated to be a marker of centrioles, of cell cycle, and of actively proliferating cells. Herein we show that this toxin at non-toxic concentrations is also capable of binding electrostatically to plasmid DNA forming DNA-peptide complexes whose stabilities overcome the need for chemical conjugation for carrying nucleic acids into cells. Interestingly, crotamine demonstrates cell specificity and targeted delivery of plasmid DNA into actively proliferating cells both in vitro and in vivo, which distinguishes crotamine from other known natural cell-penetrating peptides. The mechanism of crotamine penetration and cargo delivery into cells was also investigated, showing the involvement of heparan sulfate proteoglycans in the uptake phase, which is followed by endocytosis and peptide accumulation within the acidic endosomal vesicles. Finally, the permeabilization of endosomal membranes induced by crotamine results in the leakage of the vesicles contents to the cell cytosol.

show abstract

Section: Resultsmentioning

confidence: 99%

“…The surface electrostatic potential of crotamine is characterized by a net charge of ϩ10 (19,20). The overall structural similarities between crotamine and other natural (8,9,12,13) or synthetic, highly cationic poly-lysine-containing CPPs (21,22) encouraged us to evaluate the ability of crotamine to function as a gene delivery system.…”

mentioning

confidence: 99%

Crotamine Mediates Gene Delivery into Cells through the Binding to Heparan Sulfate Proteoglycans

Nascimento

Hayashi

Kerkis

et al. 2007

Journal of Biological Chemistry

Self Cite

104

119

View full text Add to dashboard Cite

show abstract

“…These results confirm a new cystine pairing for scorpion toxins when compared with the conventional pairs of CS␣/␤ scorpion venom peptides (C1-C4, C2-C5, and C3-C6 (11)). To further support the assigned disulfide bonds, three CYANA calculations were performed with different input parameters following a protocol described by Fadel et al (30). The first calculation included data without disulfide bond restrictions.…”

Section: For [Mϩh]mentioning

confidence: 99%

New Tricks of an Old Pattern

Saucedo

Flores-Solis

Vega

et al. 2012

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…More importantly, crotamine is also structurally related to mammalian b-defensins by forming 2-3 stranded antiparallel b-sheets with or without an a-helix at the amino-terminus. 6,8 Because of structural similarity with b-defensins, it is conceivable that crotamine might function as an AMP, in addition to being a myotoxin. Consistent with this prediction, three crotalic venoms were recently found to possess the anti-leishmanial activity, and crotamine was among the most active fractions with 50% killing of the promastigote forms of Leishmania amazonensis at approximately 20 mg ml À1 .…”

Section: Introductionmentioning

confidence: 99%

In vitro antibacterial and hemolytic activities of crotamine, a small basic myotoxin from rattlesnake Crotalus durissus

et al. 2011

View full text Add to dashboard Cite

Crotamine, a myotoxin from the venom of South American rattlesnake, is structurally related to b-defensins, antimicrobial peptides (AMPs) found in vertebrate animals. Here, we tested the antibacterial properties of crotamine and found that it killed several strains of Escherichia coli, with the MICs ranging from 25 to 100 lg ml À1 . Time-kill and bacterial membrane permeabilization assays revealed that killing of bacteria by crotamine occurred within 1 h and reached the maximum by 2 h. Additionally, the anti-E. coli activity of crotamine was completely abolished with 12.5 mM NaCl. Furthermore, the three intramolecular disulfide bonds of crotamine appeared dispensable for its antibacterial activity. The reduced form of crotamine was active against E. coli as well. However, crotamine showed no or weak activity up to 200 lg ml À1 against other species of Gram-negative and Gram-positive bacteria. Crotamine showed no appreciable hemolytic activity to erythrocytes. Our studies revealed that crotamine is also an AMP that kills bacteria through membrane permeabilization. However, crotamine appears to have a narrow antibacterial spectrum, distinct from many classical b-defensins, reinforcing the notion that crotamine originated from the b-defensin gene lineage, but has undergone significant functional diversification.

show abstract

Automated NMR structure determination and disulfide bond identification of the myotoxin crotamine from Crotalus durissus terrificus

Cited by 82 publications

References 32 publications

Crotamine Mediates Gene Delivery into Cells through the Binding to Heparan Sulfate Proteoglycans

Crotamine Mediates Gene Delivery into Cells through the Binding to Heparan Sulfate Proteoglycans

New Tricks of an Old Pattern

In vitro antibacterial and hemolytic activities of crotamine, a small basic myotoxin from rattlesnake Crotalus durissus

Contact Info

Product

Resources

About