Anti-Coagulant and Antimicrobial Recombinant Heparin-Binding Major Ampullate Spidroin 2 (MaSp2) Silk Protein.

Anti-Coagulant and Antimicrobial Recombinant Heparin-Binding Major Ampullate Spidroin 2 (MaSp2) Silk Protein.
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抗凝血和抗菌重组肝素结合大壶腹蛛丝蛋白 2 (MaSp2) 丝蛋白。

DOI:
10.3390/bioengineering9020046
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发表时间:
2022-01-19
期刊:
Bioengineering (Basel, Switzerland)
影响因子:
--
通讯作者:
Brooks AE
Brooks AE
中科院分区:
其他
文献类型:
--
作者:
Mulinti P;Diekjürgen D;Kurtzeborn K;Balasubramanian N;Stafslien SJ;Grainger DW;Brooks AE

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在已建立的结构-性质关系的支配下,由主要壶腹蜘蛛丝组成的多肽主题提供了强度和延展性的平衡。与特定功能相关的其他受生物启发的小肽基序可以在这些单元中组合,以创造具有新杂交特性的设计师丝绸材料。在这项研究中,一种已知既能结合肝素又能模拟抗菌肽的小碱性多肽(ARKKAAKA)与一种抗蛋白酶、机械强度高的丝状多肽MaSp2有基因联系。纯化的融合蛋白(4个丝素结构域和4个肝素结合肽重复序列)在大肠杆菌中得到了表达。多种分析分析表明,MaSp2蜘蛛丝肽与肝素结合基序成功融合。用ELISA法检测融合多肽与肝素结合的能力,并用αPTT法进一步检测其抗凝血活性。采用抑菌圈法和结晶紫(CV)法对其抑菌性能进行了评价。使用这一策略,我们能够将两种类型的遗传基序联系起来,创造出一种设计的丝状蛋白,具有更好的血液相容性和抗菌性能。
Governed by established structure–property relationships, peptide motifs comprising major ampullate spider silk confer a balance of strength and extensibility. Other biologically inspired small peptide motifs correlated to specific functionalities can be combined within these units to create designer silk materials with new hybrid properties. In this study, a small basic peptide, (ARKKAAKA) known to both bind heparin and mimic an antimicrobial peptide, was genetically linked to a protease-resistant, mechanically robust silk-like peptide, MaSp2. Purified fusion proteins (four silk domains and four heparin-binding peptide repeats) were expressed in E. coli. Successful fusion of a MaSp2 spider silk peptide with the heparin-binding motif was shown using a variety of analytical assays. The ability of the fusion peptide to bind heparin was assessed with ELISA and was further tested for its anticoagulant property using aPTT assay. Its intrinsic property to inhibit bacterial growth was evaluated using zone of inhibition and crystal violet (CV) assays. Using this strategy, we were able to link the two types of genetic motifs to create a designer silk-like protein with improved hemocompatibility and antimicrobial properties.
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