A new temporin with antibacterial activity and cytotoxicity from the skin secretion of Lithobates palmipes (Spix, 1824) (Amphibia: Ranidae) from Brazilian Atlantic Forest

IF 1.9 3区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology Pub Date : 2024-10-18 DOI:10.1016/j.cbpb.2024.111041

Géssica Gomes Barbosa , Carlos José Correia de Santana , Tulíbia Laurindo Silva , Brenda Conceição Guimarães Santana , Patrícia Maria Guedes Paiva , Gabriel Gonçalves de Freitas , Guilherme Dotto Brand , Osmindo Rodrigues Pires Júnior , Mariana S. Castro , Thiago Henrique Napoleão

{"title":"A new temporin with antibacterial activity and cytotoxicity from the skin secretion of Lithobates palmipes (Spix, 1824) (Amphibia: Ranidae) from Brazilian Atlantic Forest","authors":"Géssica Gomes Barbosa , Carlos José Correia de Santana , Tulíbia Laurindo Silva , Brenda Conceição Guimarães Santana , Patrícia Maria Guedes Paiva , Gabriel Gonçalves de Freitas , Guilherme Dotto Brand , Osmindo Rodrigues Pires Júnior , Mariana S. Castro , Thiago Henrique Napoleão","doi":"10.1016/j.cbpb.2024.111041","DOIUrl":null,"url":null,"abstract":"<div><div>This work investigated the peptide profile of skin secretion from Lithobates palmipes collected from the Brazilian Atlantic Forest. The secretion was submitted to reversed phase high-performance liquid chromatography (RP-HPLC) and the fractions were screened for antibacterial activity. RP-HPLC resulted in the separation of several peaks, among which 10 showed antibacterial activity and contained peptides of the ranatuerin, brevinin and temporin families. Fraction 6 was resubmitted to RP-HPLC and a novel peptide from temporin family (temporin-PMb) had its primary structure determined. Temporin-PMb and non-amidated temporin-PMb were synthesized, purified, and evaluated for antibacterial activity, hemolytic activity and cytotoxicity to keratinocytes and cancer cells. Temporin-PMb was active against Klebsiella pneumoniae, Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa as well as against methicilin-resistant S. aureus (MRSA) and Acinetobacter baumannii. It was cytotoxic to human cervical adenocarcinoma cells (HeLa) and human mammary adenocarcinoma cells (MCF7) with IC50 of 32.4 and 24.1 μM, respectively. It was also toxic to human keratinocytes (HaCaT; IC50 of 25.0 μM) and showed hemolytic activity. The non-amidated form showed low hemolytic activity and lower HaCaT toxicity, but was only effective against E. coli, S. aureus MRSA, and A. baumanii. In conclusion, Atlantic Forest L. palmipes skin secretion contained different bioactive peptides, including a novel temporin with antibacterial effect and cytotoxicity towards human cancer cells. The amide group was responsible for the activities of the wild-type temporin-PMb. Peptide engineering studies are encouraged aiming at minimizing unwanted effects.</div></div>","PeriodicalId":55236,"journal":{"name":"Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology","volume":"275 ","pages":"Article 111041"},"PeriodicalIF":1.9000,"publicationDate":"2024-10-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology","FirstCategoryId":"99","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1096495924001088","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}

引用次数: 0

Abstract

This work investigated the peptide profile of skin secretion from Lithobates palmipes collected from the Brazilian Atlantic Forest. The secretion was submitted to reversed phase high-performance liquid chromatography (RP-HPLC) and the fractions were screened for antibacterial activity. RP-HPLC resulted in the separation of several peaks, among which 10 showed antibacterial activity and contained peptides of the ranatuerin, brevinin and temporin families. Fraction 6 was resubmitted to RP-HPLC and a novel peptide from temporin family (temporin-PMb) had its primary structure determined. Temporin-PMb and non-amidated temporin-PMb were synthesized, purified, and evaluated for antibacterial activity, hemolytic activity and cytotoxicity to keratinocytes and cancer cells. Temporin-PMb was active against Klebsiella pneumoniae, Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa as well as against methicilin-resistant S. aureus (MRSA) and Acinetobacter baumannii. It was cytotoxic to human cervical adenocarcinoma cells (HeLa) and human mammary adenocarcinoma cells (MCF7) with IC₅₀ of 32.4 and 24.1 μM, respectively. It was also toxic to human keratinocytes (HaCaT; IC₅₀ of 25.0 μM) and showed hemolytic activity. The non-amidated form showed low hemolytic activity and lower HaCaT toxicity, but was only effective against E. coli, S. aureus MRSA, and A. baumanii. In conclusion, Atlantic Forest L. palmipes skin secretion contained different bioactive peptides, including a novel temporin with antibacterial effect and cytotoxicity towards human cancer cells. The amide group was responsible for the activities of the wild-type temporin-PMb. Peptide engineering studies are encouraged aiming at minimizing unwanted effects.

Abstract Image

查看原文本刊更多论文

来自巴西大西洋森林的 Lithobates palmipes (Spix, 1824) (Amphibia: Ranidae) 皮肤分泌物的一种具有抗菌活性和细胞毒性的新颞素。

这项研究调查了从巴西大西洋森林采集的棕榈栗蝠（Lithobates palmipes）皮肤分泌物的肽谱。分泌物被送入反相高效液相色谱法（RP-HPLC），并对馏分进行抗菌活性筛选。反相高效液相色谱法分离出几个峰值，其中 10 个峰值具有抗菌活性，含有 ranatuerin、brevinin 和 temporin 家族的肽。对馏分 6 再次进行了 RP-HPLC 分析，并确定了一种来自颞素家族的新型多肽（颞素-PMb）的一级结构。对 Temporin-PMb 和非酰胺化的 temporin-PMb 进行了合成、纯化，并对其抗菌活性、溶血活性以及对角质细胞和癌细胞的细胞毒性进行了评估。Temporin-PMb 对肺炎克雷伯氏菌、金黄色葡萄球菌、大肠埃希菌和铜绿假单胞菌以及耐甲氧西林金黄色葡萄球菌（MRSA）和鲍曼不动杆菌具有活性。它对人宫颈腺癌细胞（HeLa）和人乳腺腺癌细胞（MCF7）具有细胞毒性，IC50 分别为 32.4 和 24.1 μM。它对人类角质细胞（HaCaT；IC50 为 25.0 μM）也有毒性，并显示出溶血活性。非酰胺化形式的溶血活性较低，对 HaCaT 的毒性也较低，但只对大肠杆菌、金黄色葡萄球菌 MRSA 和鲍曼不动杆菌有效。总之，大西洋森林 L. palmipes 皮肤分泌物含有不同的生物活性肽，其中包括一种具有抗菌作用和对人类癌细胞细胞毒性的新型颞素。酰胺基团是野生型颞素-PMb活性的来源。我们鼓励进行肽工程研究，以尽量减少不必要的影响。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Comparative Biochemistry and Physiology B-Biochemistry & Molecular Biology 生物-动物学

CiteScore

4.60

自引率

4.50%

发文量

审稿时长

22 days

期刊介绍： Comparative Biochemistry & Physiology (CBP) publishes papers in comparative, environmental and evolutionary physiology. Part B: Biochemical and Molecular Biology (CBPB), focuses on biochemical physiology, primarily bioenergetics/energy metabolism, cell biology, cellular stress responses, enzymology, intermediary metabolism, macromolecular structure and function, gene regulation, evolutionary genetics. Most studies focus on biochemical or molecular analyses that have clear ramifications for physiological processes.