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基于人弹性蛋白样多肽(HELPs)模型系统的仿生 3D 基质用于弹性酶活性检测的评估。

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

机构信息

Department of Life Sciences, University of Trieste, 34127, Trieste, Italy.

Microbiology Unit, University Hospital ASUITS, 34149, Trieste, Italy.

出版信息

J Biotechnol. 2017 Aug 10;255:57-65. doi: 10.1016/j.jbiotec.2017.06.006. Epub 2017 Jun 15.

Abstract

Elastin is a fibrous protein that confers elasticity to tissues such as skin, arteries and lung. It is extensively cross-linked, 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.

摘要

弹性蛋白是一种纤维状蛋白质,赋予皮肤、动脉和肺等组织弹性。它广泛交联,高度疏水且不溶。然而,弹性蛋白可以在感染性疾病中被细菌蛋白酶水解,导致或多或少严重的组织损伤。因此,需要开发能够可靠且特异性检测病原体分泌的弹性蛋白水解活性的底物,以改善对细菌蛋白酶可能引起的损伤的体外评估。在这项工作中,两种人类仿生弹性蛋白多肽,HELP 和 HELP1,以及源自 HELP 的基质,被用作弹性蛋白水解活性检测的底物。从囊性纤维化患者中分离的 30 株铜绿假单胞菌与标准底物平行分析,以检测蛋白水解和弹性蛋白水解活性。结果表明,基于 HELP 的 3D 基质是一种有前途的弹性蛋白仿生模型,可用于体外评估细菌弹性蛋白水解活性。此外,这种模型底物还可以进一步阐明分子水平上弹性蛋白降解的机制,并开发对外部刺激有响应的基于仿生材料的装置。

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