Volume 40 Issue 3
May  2019
Turn off MathJax
Article Contents

Fen Zhang, Zhi-Lai Guo, Yan Chen, Li Li, Hai-Ning Yu, Yi-Peng Wang. Effects of C-terminal amidation and heptapeptide ring on the biological activities and advanced structure of amurin-9KY, a novel antimicrobial peptide identified from the brown frog, Rana kunyuensis[J]. Zoological Research, 2019, 40(3): 198-204. doi: 10.24272/j.issn.2095-8137.2018.070
Citation: Fen Zhang, Zhi-Lai Guo, Yan Chen, Li Li, Hai-Ning Yu, Yi-Peng Wang. Effects of C-terminal amidation and heptapeptide ring on the biological activities and advanced structure of amurin-9KY, a novel antimicrobial peptide identified from the brown frog, Rana kunyuensis[J]. Zoological Research, 2019, 40(3): 198-204. doi: 10.24272/j.issn.2095-8137.2018.070

Effects of C-terminal amidation and heptapeptide ring on the biological activities and advanced structure of amurin-9KY, a novel antimicrobial peptide identified from the brown frog, Rana kunyuensis

doi: 10.24272/j.issn.2095-8137.2018.070
Funds:  This work was supported by grants from the National Natural Science Foundation of China (31772455), Natural Science Foundation of Jiangsu Province (BK20160336 and BK20171214), Natural Science Foundation of College in Jiangsu Province (16KJB350004), Suzhou Science and Technology Development Project (SYN201504 and SNG2017045)
More Information
  • Corresponding author: Hai-Ning Yu, Yi-Peng Wang
  • Received Date: 2018-06-20
  • Publish Date: 2019-05-18
  • Rana kunyuensis is a species of brown frog that lives exclusively on Kunyu Mountain, Yantai, China. In the current study, a 279-bp cDNA sequence encoding a novel antimicrobial peptide (AMP), designated as amurin-9KY, was cloned from synthesized double-strand skin cDNA of R. kunyuensis. The amurin-9KY precursor was composed of 62 amino acid (aa) residues, whereas the mature peptide was composed of 14 aa and contained two cysteines forming a C-terminal heptapeptide ring (Rana box domain) and an amidated C-terminus. These structural characters represent a novel amphibian AMP family. Although amurin-9KY exhibited high similarity to the already identified amurin-9AM from R. amurensis, little is known about the structures and activities of amurin-9 family AMPs so far. Therefore, amurin-9KY and its three derivatives (amurin-9KY1–3) were designed and synthesized. The structures and activities were examined to evaluate the influence of C-terminal amidation and the heptapeptide ring on the activities and structure of amurin-9KY. Results indicated that C-terminal amidation was essential for antimicrobial activity, whereas both C-terminal amidation and the heptapeptide ring played roles in the low hemolytic activity. Circular dichroism (CD) spectra showed that the four peptides adopted an a-helical conformation in THF/H2O (v/v 1:1) solution, but a random coil in aqueous solution. Elimination of the C-terminal heptapeptide ring generated two free cysteine residues with unpaired thiol groups, which greatly increased the concentration-dependent anti-oxidant activity. Scanning electron microscopy (SEM) was also performed to determine the possible bactericidal mechanisms.
  • 加载中
  • 加载中
通讯作者: 陈斌, bchen63@163.com
  • 1. 

    沈阳化工大学材料科学与工程学院 沈阳 110142

  1. 本站搜索
  2. 百度学术搜索
  3. 万方数据库搜索
  4. CNKI搜索

Article Metrics

Article views(635) PDF downloads(191) Cited by()

Related
Proportional views

Effects of C-terminal amidation and heptapeptide ring on the biological activities and advanced structure of amurin-9KY, a novel antimicrobial peptide identified from the brown frog, Rana kunyuensis

doi: 10.24272/j.issn.2095-8137.2018.070
Funds:  This work was supported by grants from the National Natural Science Foundation of China (31772455), Natural Science Foundation of Jiangsu Province (BK20160336 and BK20171214), Natural Science Foundation of College in Jiangsu Province (16KJB350004), Suzhou Science and Technology Development Project (SYN201504 and SNG2017045)
    Corresponding author: Hai-Ning Yu, Yi-Peng Wang

Abstract: Rana kunyuensis is a species of brown frog that lives exclusively on Kunyu Mountain, Yantai, China. In the current study, a 279-bp cDNA sequence encoding a novel antimicrobial peptide (AMP), designated as amurin-9KY, was cloned from synthesized double-strand skin cDNA of R. kunyuensis. The amurin-9KY precursor was composed of 62 amino acid (aa) residues, whereas the mature peptide was composed of 14 aa and contained two cysteines forming a C-terminal heptapeptide ring (Rana box domain) and an amidated C-terminus. These structural characters represent a novel amphibian AMP family. Although amurin-9KY exhibited high similarity to the already identified amurin-9AM from R. amurensis, little is known about the structures and activities of amurin-9 family AMPs so far. Therefore, amurin-9KY and its three derivatives (amurin-9KY1–3) were designed and synthesized. The structures and activities were examined to evaluate the influence of C-terminal amidation and the heptapeptide ring on the activities and structure of amurin-9KY. Results indicated that C-terminal amidation was essential for antimicrobial activity, whereas both C-terminal amidation and the heptapeptide ring played roles in the low hemolytic activity. Circular dichroism (CD) spectra showed that the four peptides adopted an a-helical conformation in THF/H2O (v/v 1:1) solution, but a random coil in aqueous solution. Elimination of the C-terminal heptapeptide ring generated two free cysteine residues with unpaired thiol groups, which greatly increased the concentration-dependent anti-oxidant activity. Scanning electron microscopy (SEM) was also performed to determine the possible bactericidal mechanisms.

Fen Zhang, Zhi-Lai Guo, Yan Chen, Li Li, Hai-Ning Yu, Yi-Peng Wang. Effects of C-terminal amidation and heptapeptide ring on the biological activities and advanced structure of amurin-9KY, a novel antimicrobial peptide identified from the brown frog, Rana kunyuensis[J]. Zoological Research, 2019, 40(3): 198-204. doi: 10.24272/j.issn.2095-8137.2018.070
Citation: Fen Zhang, Zhi-Lai Guo, Yan Chen, Li Li, Hai-Ning Yu, Yi-Peng Wang. Effects of C-terminal amidation and heptapeptide ring on the biological activities and advanced structure of amurin-9KY, a novel antimicrobial peptide identified from the brown frog, Rana kunyuensis[J]. Zoological Research, 2019, 40(3): 198-204. doi: 10.24272/j.issn.2095-8137.2018.070

Catalog

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return