Bombinin-like peptides with antimicrobial activity from skin secretions of the Asian toad, Bombina orientalis.
Bombinin-like peptides with antimicrobial activity from skin secretions of the Asian toad, Bombina orientalis.
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DOI：
10.1016/s0021-9258(18)54469-0
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发表时间：
1991-12
期刊：
影响因子：
--
通讯作者：
B. Gibson;D. Tang;R. Mandrell;M. Kelly;E. Spindel
中科院分区：
文献类型：
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作者：
B. Gibson;D. Tang;R. Mandrell;M. Kelly;E. Spindel
本文报道了从东方铃蟾（Bombina orientalis）皮肤中分离得到的三种铃蟾肽（BLP-1-3）的结构和溶血、杀菌活性。从B的皮肤中分离肽。orientalis并通过串联质谱法测序，并且是长度为25-27个氨基酸的两亲性阳离子肽。最丰富的成员（BLP-1）的序列是：Gly-Ile-Gly-Ala-Ser-Ile-Leu-Ser-Ala-Gly-Lys-Ser-Ala-Leu-Lys-Gly-Leu-Ala-Lys-Gly-Leu-Ala-Glu-His-Phe-Ala-Asn-NH 2。发现所有三种肽与铃蟾肽（bombinin）具有相当大的但不完全的同源性，铃蟾肽是Michl和Csordas首先分离的抗微生物溶血肽（Csordas，A.，和Michl，A. 03 The Dog of the Woman（1970）101，182-189）从铃蟾（Bombina variegata）的皮肤中分离。BLP已经被测定了抗生素和溶血活性，并且发现在它们杀死细菌的能力方面比爪蟾抗菌肽2（来自非洲爪蟾的相关抗微生物肽）更有效。然而，没有发现这些肽的显着溶血活性，这表明对原核生物膜的选择性超过真核生物膜。抗菌活性的分子基础被认为是由于其预测的两亲性α-螺旋结构，这得到了圆二色性测量的支持，圆二色性测量在40%三氟乙醇中发现了显著的螺旋含量（63-69% α-螺旋）。最后，构建了B皮肤cDNA文库。orientalis中，并用与BLP-1的COOH末端互补的寡核苷酸探针进行筛选。分离并测序了编码BLP-1和BLP-3以及与BLP-3相差两个氨基酸取代的另外的肽（BLP-4）的几个克隆。
The structures and hemolytic and bactericidal activities of three bombinin-like peptides, or BLP-1-3, from the skin of Bombina orientalis are described. The peptides were isolated from the skin of B. orientalis and sequenced by tandem mass spectrometry and are amphipathic, cationic peptides of 25-27 amino acids in length. The sequence of the most abundant member (BLP-1) is: Gly-Ile-Gly-Ala-Ser-Ile-Leu-Ser-Ala-Gly-Lys-Ser-Ala-Leu-Lys-Gly-Leu- Ala-Lys-Gly-Leu-Ala-Glu-His-Phe-Ala-Asn-NH2. All three peptides were found to share considerable, but not complete, homology with bombinin, an antimicrobial, hemolytic peptide first isolated by Michl and Csordas (Csordas, A., and Michl, A. (1970) Monatsh. Chem. 101, 182-189) from the skin of Bombina variegata. The BLPs have been assayed for antibiotic and hemolytic activity and found to be more potent than magainin 2 (a related antimicrobial peptide from Xenopus laevis) in their ability to kill bacteria. However, no significant hemolytic activity was found for these peptides which suggests a selectivity for prokaryotic over eukaryotic membranes. The molecular basis for antibacterial activity is presumed to be due to their predicted amphipathic alpha-helical structures which is supported by circular dichroism measurements that found significant helical content (63-69% alpha-helix) in 40% trifluoroethanol. Last, a cDNA library was constructed from the skin of B. orientalis and screened with an oligonucleotide probe complementary to the COOH terminus of BLP-1. Several clones were isolated and sequenced that encode BLP-1 and BLP-3, as well as an additional peptide (BLP-4) that differs by two amino acid substitutions from BLP-3.