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首页> 外文期刊>World Journal of Microbiology & Biotechnology >Purification, molecular cloning, and antimicrobial activity of peptides from the skin secretion of the black-spotted frog, Rana nigromaculata
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Purification, molecular cloning, and antimicrobial activity of peptides from the skin secretion of the black-spotted frog, Rana nigromaculata

机译：黑斑蛙黑蛙蛙皮肤分泌物中多肽的纯化，分子克隆和抗菌活性

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摘要

Antimicrobial peptides from a wide range of amphibian species, especially frogs of the genus Rana, have been characterised and are potential therapeutic agents. Here we describe the isolation, purification, and structural and biological characterisation of three novel antimicrobial peptides from the skin secretions of the black spotted frog, Rana nigromaculata, from Northeastern China. The peptides were identified as belonging to two known families: the temporin, which was first identified in R. nigromaculata from China, and the brevinin-2. Temporin-1RNa and temporin-1RNb both containing three positive charges and have a high potency against microorganisms (MIC: 3.13-8.3 mu M against Gram-positive bacteria, 12.5-25.0 mu M against Gram-negative bacteria, and 6.25-12.5 mu M against Candida albicans) and a high haemolytic activity against human erythrocytes (HC50: 100-150 mu M). Brevinin-2RNa contains a single intra-disulphide bridge at the C-terminus that is active towards the tested Gram-positive bacteria but is not active against E. coli and P. aeruginosa. The cDNAs encoding three novel peptide precursors were also subsequently cloned from an R. nigromaculata skin cDNA library and sequenced. The precursors contain 58-72 amino acid residues, which include a conserved signal peptide, acidic propeptide, and the mature temporin-1RNa, temporin-1RNb and brevinin-2RNa. The CD spectra of temporin-1RNa and temporin-1RNb in water, 30 mM SDS and 50 % trifluoroethanol (TFE) indicated that both peptides adopted an aperiodic structure in water and an organised structure with an alpha-helical conformation in TFE and SDS solution. The conformational transition induced by TFE or SDS reflects the potential ability of temporin-1RNa and temporin-1RNb to interact with anionic membranes.
机译：来自多种两栖动物，尤其是蛙属蛙的抗菌肽已被表征，并且是潜在的治疗剂。在这里，我们从中国东北黑斑蛙蛙皮肤黑蛙的皮肤分泌物中描述了三种新型抗菌肽的分离，纯化以及结构和生物学特性。这些肽被鉴定为属于两个已知家族：temporin和brevinin-2，temporin是最早在中国的黑松鼠（R. nigromaculata）中鉴定的。 Temporin-1RNa和temporin-1RNb都含有三个正电荷，并且对微生物具有很高的效价（MIC：对革兰氏阳性菌为3.13-8.3μM，对革兰氏阴性菌为12.5-25.0μM，以及6.25-12.5μM抗白色念珠菌）和高抗人红细胞溶血活性（HC50：100-150μM）。 Brevinin-2RNa在C端包含一个二硫键，对被测革兰氏阳性细菌有活性，但对大肠杆菌和铜绿假单胞菌没有活性。随后，还从黑麦草皮肤皮肤cDNA文库中克隆了编码三种新型肽前体的cDNA并进行了测序。前体含有58-72个氨基酸残基，其中包括一个保守的信号肽，酸性肽和成熟的Temporin-1RNa，temporin-1RNb和brevinin-2RNa。 temporin-1RNa和temporin-1RNb在水中，30 mM SDS和50％三氟乙醇（TFE）中的CD光谱表明，这两种肽在水中均采用非周期性结构，并在TFE和SDS溶液中采用α-螺旋构象的有序结构。 TFE或SDS诱导的构象转变反映了Temporin-1RNa和temporin-1RNb与阴离子膜相互作用的潜在能力。

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