Evaluation of a biomimetic 3D substrate based on the Human Elastin-like Polypeptides (HELPs) model system for elastolytic activity detection

被引:13
作者
Corich, Lucia [1 ]
Busetti, Marina [2 ]
Petix, Vincenzo [3 ]
Passamonti, Sabina [1 ]
Bandiera, Antonella [1 ]
机构
[1] Univ Trieste, Dept Life Sci, I-34127 Trieste, Italy
[2] Univ Hosp ASUITS, Microbiol Unit, I-34149 Trieste, Italy
[3] IRCCS Burlo Garofolo, Inst Maternal & Child Hlth, I-34137 Trieste, Italy
关键词
Biomimetic substrate; 3D matrix; elastin; elastolytic activity assay; PSEUDOMONAS-AERUGINOSA STRAINS; MATRIX METALLOPROTEINASES; DEGRADATION; SPECIFICITY; HYDROLYSIS; PROTEASE; LASA;
D O I
10.1016/j.jbiotec.2017.06.006
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Elastin is a fibrous protein that confers elasticity to tissues such as skin, arteries and lung. It is extensively crosslinked, highly hydrophobic and insoluble. Nevertheless, elastin can be hydrolysed by bacterial proteases in infectious diseases, resulting in more or less severe tissue damage. Thus, development of substrates able to reliably and specifically detect pathogen-secreted elastolytic activity is needed to improve the in vitro evaluation of the injury that bacterial proteases may provoke. In this work, two human biomimetic elastin polypeptides, HELP and HELP1, as well as the matrices derived from HELP, have been probed as substrates for elastolytic activity detection. Thirty strains of Pseudomonas aeruginosa isolated from cystic fibrosis patients were analyzed in parallel with standard substrates, to detect proteolytic and elastolytic activity. Results point to the HELP-based 3D matrix as an interesting biomimetic model of elastin to assess bacterial elastolytic activity in vitro. Moreover, this model substrate enables to further elucidate the mechanism underlying elastin degradation at molecular level, as well as to develop biomimetic material-based devices responsive to external stimuli.
引用
收藏
页码:57 / 65
页数:9
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