Antimicrobial peptide from the skin secretion of the frog Leptodactylus syphax

Dourado, Flávio Santos; Leite, José Roberto S. A.; Silva, Luciano; Melo, Jorge Alex Taquita; Bloch, Carlos; Schwartz, Elisabeth Ferroni

doi:10.1016/j.toxicon.2007.04.027

Cited by 39 publications

(32 citation statements)

References 28 publications

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“…These sequences were aligned with Clustal Omega [52]. Sequences are from the following references: ocellatin-S1 [23], ocellatin-1 [53], ocellatin-K1 and ocellatin-PT3 [20] …”

Section: Resultsmentioning

confidence: 99%

Ocellatin peptides from the skin secretion of the South American frog Leptodactylus labyrinthicus (Leptodactylidae): characterization, antimicrobial activities and membrane interactions

Gusmao

Santos

et al. 2017

J Venom Anim Toxins Incl Trop Dis

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BackgroundThe availability of antimicrobial peptides from several different natural sources has opened an avenue for the discovery of new biologically active molecules. To the best of our knowledge, only two peptides isolated from the frog Leptodactylus labyrinthicus, namely pentadactylin and ocellatin-F1, have shown antimicrobial activities. Therefore, in order to explore the antimicrobial potential of this species, we have investigated the biological activities and membrane interactions of three peptides isolated from the anuran skin secretion.MethodsThree peptide primary structures were determined by automated Edman degradation. These sequences were prepared by solid-phase synthesis and submitted to activity assays against gram-positive and gram-negative bacteria and against two fungal strains. The hemolytic properties of the peptides were also investigated in assays with rabbit blood erythrocytes. The conformational preferences of the peptides and their membrane interactions have been investigated by circular dichroism spectroscopy and liposome dye release assays.ResultsThe amino acid compositions of three ocellatins were determined and the sequences exhibit 100% homology for the first 22 residues (ocellatin-LB1 sequence). Ocellatin-LB2 carries an extra Asn residue and ocellatin-F1 extra Asn-Lys-Leu residues at C-terminus. Ocellatin-F1 presents a stronger antibiotic potential and a broader spectrum of activities compared to the other peptides. The membrane interactions and pore formation capacities of the peptides correlate directly with their antimicrobial activities, i.e., ocellatin-F1 > ocellatin-LB1 > ocellatin-LB2. All peptides acquire high helical contents in membrane environments. However, ocellatin-F1 shows in average stronger helical propensities.ConclusionsThe obtained results indicate that the three extra amino acid residues at the ocellatin-F1 C-terminus play an important role in promoting stronger peptide-membrane interactions and antimicrobial properties. The extra Asn-23 residue present in ocellatin-LB2 sequence seems to decrease its antimicrobial potential and the strength of the peptide-membrane interactions.Electronic supplementary materialThe online version of this article (doi:10.1186/s40409-017-0094-y) contains supplementary material, which is available to authorized users.

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“…These sequences were aligned with Clustal Omega [52]. Sequences are from the following references: ocellatin-S1 [23], ocellatin-1 [53], ocellatin-K1 and ocellatin-PT3 [20] …”

Section: Resultsmentioning

confidence: 99%

Ocellatin peptides from the skin secretion of the South American frog Leptodactylus labyrinthicus (Leptodactylidae): characterization, antimicrobial activities and membrane interactions

Gusmao

Santos

et al. 2017

J Venom Anim Toxins Incl Trop Dis

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“…fallax, Krintler, 1986), and the dermal glands of several species are known to produce defensive skin secretions (Cei et al, 1967), including antimicrobial peptides and other protein toxins found in L. fallax (Rollins-Smith et al, 2005;King et al, 2005a), L. labyrinthicus (Libério et al, 2014), L. laticeps (Conlon et al, 2009), L. latrans (Nascimento et al, 2004(Nascimento et al, , 2007Leite et al, 2010), L. pentadactylus (Habermehl, 1981;Barlow, 1998;King et al, 2005b;Limaverde et al, 2009;Sousa et al, 2009), L. syphax (Dourado et al, 2007). Similarly, the foam nests of L. vastus possess a novel surfactant protein (Hissa et al, 2008(Hissa et al, , 2014.…”

Section: Introductionmentioning

confidence: 99%

Systematics of the Neotropical GenusLeptodactylusFitzinger, 1826 (Anura: Leptodactylidae): Phylogeny, the Relevance of Non-molecular Evidence, and Species Accounts

Sá

Grant

Camargo

et al. 2014

South American Journal of Herpetology

130

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“…Leite et al [26] identified five truncated forms of ocellatins from L. ocellatus . Dourado et al [24] found six fragments of the peptide syphaxin from L. syphax . In addition, a 22-residue fallaxin fragment identified by our group was isolated by Rollins-Smith et al [22] from the skin secretion of L. fallax as well.…”

Section: Resultsmentioning

confidence: 99%

“…In this way, the antimicrobial activity lost with hydrolysis can be restored [65], or the peptides may acquire a new function. On the other hand, there are reports regarding peptide fragments such as SPX(1–16) and SPX(1–22), derived from the AMP syphaxin, which display potent biological activity without any processing after hydrolysis [24].…”

Section: Discussionmentioning

confidence: 99%

“…Previously, we detected two inactive fragments of the AMP fallaxin in the skin secretion of Leptodactylus labyrinthicus , which called our attention to the enzymes involved in these proteolytic cleavages. Other research groups also have identified the presence of AMP fragments in their studies [22], [24], [26]. Therefore, the aim of the present study was to identify and characterize the proteolytic activity present in the crude skin secretion of the pepper frog L. labyrinthicus .…”

Section: Introductionmentioning

confidence: 93%

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The Crude Skin Secretion of the Pepper Frog Leptodactylus labyrinthicus Is Rich in Metallo and Serine Peptidases

et al. 2014

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Peptidases are ubiquitous enzymes involved in diverse biological processes. Fragments from bioactive peptides have been found in skin secretions from frogs, and their presence suggests processing by peptidases. Thus, the aim of this work was to characterize the peptidase activity present in the skin secretion of Leptodactylus labyrinthicus. Zymography revealed the presence of three bands of gelatinase activity of approximately 60 kDa, 66 kDa, and 80 kDa, which the first two were calcium-dependent. These three bands were inhibited either by ethylenediaminetetraacetic acid (EDTA) and phenathroline; thus, they were characterized as metallopeptidases. Furthermore, the proteolytic enzymes identified were active only at pH 6.0–10.0, and their activity increased in the presence of CHAPS or NaCl. Experiments with fluorogenic substrates incubated with skin secretions identified aminopeptidase activity, with cleavage after leucine, proline, and alanine residues. This activity was directly proportional to the protein concentration, and it was inhibited in the presence of metallo and serine peptidase inhibitors. Besides, the optimal pH for substrate cleavage was determined to be 7.0–8.0. The results of the in gel activity assay showed that all substrates were hydrolyzed by a 45 kDa peptidase. Gly-Pro-AMC was also cleaved by a peptidase greater than 97 kDa. The data suggest the presence of dipeptidyl peptidases (DPPs) and metallopeptidases; however, further research is necessary. In conclusion, our work will help to elucidate the implication of these enzymatic activities in the processing of the bioactive peptides present in frog venom, expanding the knowledge of amphibian biology.

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Antimicrobial peptide from the skin secretion of the frog Leptodactylus syphax

Cited by 39 publications

References 28 publications

Ocellatin peptides from the skin secretion of the South American frog Leptodactylus labyrinthicus (Leptodactylidae): characterization, antimicrobial activities and membrane interactions

Ocellatin peptides from the skin secretion of the South American frog Leptodactylus labyrinthicus (Leptodactylidae): characterization, antimicrobial activities and membrane interactions

Systematics of the Neotropical GenusLeptodactylusFitzinger, 1826 (Anura: Leptodactylidae): Phylogeny, the Relevance of Non-molecular Evidence, and Species Accounts

The Crude Skin Secretion of the Pepper Frog Leptodactylus labyrinthicus Is Rich in Metallo and Serine Peptidases

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