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用于 DNA 折纸涂层的聚合物用白蛋白大分子引发剂的开发。

Development of albumin macroinitiator for polymers to use in DNA origami coating.

机构信息

Department of Nanotechnology and Nanomedicine, Graduate School of Science and Engineering, Hacettepe University, Ankara, Turkey

出版信息

Turk J Med Sci. 2020 Aug 26;50(5):1461-1469. doi: 10.3906/sag-2001-245.

DOI:10.3906/sag-2001-245
PMID:32283899
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7491299/
Abstract

BACKGROUND/AIM: DNA nanostructures have many advantages over polymers and lipid based drug delivery agents such as biodegradability and biocompatibility. However their transfection rates and stability still limit their widely use in nanomedicine. In this study highly versatile and straightforward albumin coating preparation method is showed for DNA nanostructures.

MATERIALS AND METHODS

N-methylolmaleimide was esterified with a-bromoisobutyrl bromide (BiBB) to achive bromine functional structure. Then it was attached to bovine serum albumin (BSA) via cysteine-maleimide bond further to use as macroinitiator for atom transfer radical polymerization (ATRP). Cationic polymers can be synthesized from this end further to use as binding domain for fabricated 60 Helix bundle DNA origami.

RESULTS

Proton nuclear magnetic resonance (1H NMR) analysis used for characterization. Methyelene group hydrogens’ peak in 5.0 ppm and strong peak in 1.5–2.0 ppm range showed proper methylolation of maleimide and bromine functional formation, respectively. Then BSA-macroinitiator formation is verified by 1780 Da peak shift in MALDI-TOF (Matrix-assisted laser desorption/ionization - time of flight) spectrum. Moreover electrophoretic mobility shift assay (EMSA) showed successful dense 60 Helix bundle formation.

CONCLUSION

In this study, a facile method is developed to synthesize protein conjugated-ATRP initiator further can be used in polymerization and coating DNA nanostructures. It is feasible for any protein containing cysteine amino acid.

摘要

背景/目的：DNA 纳米结构在生物降解性和生物相容性方面优于聚合物和基于脂质的药物递送剂，具有许多优势。然而，它们的转染率和稳定性仍然限制了它们在纳米医学中的广泛应用。在这项研究中，展示了一种非常通用且简单的白蛋白涂层制备方法，用于 DNA 纳米结构。

材料和方法

N-羟甲基马来酰亚胺与溴异丁酰溴（BiBB）酯化，得到溴功能结构。然后通过半胱氨酸-马来酰亚胺键将其连接到牛血清白蛋白（BSA）上，进一步用作原子转移自由基聚合（ATRP）的大分子引发剂。可以从这个末端合成阳离子聚合物，用作合成的 60 螺旋束 DNA 折纸的结合域。

结果

质子核磁共振（1H NMR）分析用于表征。5.0 ppm 处亚甲基氢的峰和 1.5-2.0 ppm 范围内的强峰分别表明马来酰亚胺的适当甲氧基化和溴功能形成。然后通过 MALDI-TOF（基质辅助激光解吸/电离-飞行时间）光谱中的 1780 Da 峰位移验证 BSA-大分子引发剂的形成。此外，电泳迁移率变动分析（EMSA）显示出成功的密集 60 螺旋束形成。

结论

在这项研究中，开发了一种简便的方法来合成蛋白质偶联的 ATRP 引发剂，进一步可用于聚合和涂层 DNA 纳米结构。它适用于任何含有半胱氨酸氨基酸的蛋白质。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8023/7491299/f547e12a8fda/turkjmedsci-50-1461-fig010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8023/7491299/68a9f06c8266/turkjmedsci-50-1461-fig001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8023/7491299/c84a99e1fc87/turkjmedsci-50-1461-fig002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8023/7491299/ccbfd4f7035f/turkjmedsci-50-1461-fig003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8023/7491299/14346415dc6e/turkjmedsci-50-1461-fig004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8023/7491299/25482dbe6fbc/turkjmedsci-50-1461-fig005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8023/7491299/43a82218e497/turkjmedsci-50-1461-fig006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8023/7491299/a21be9ba08d9/turkjmedsci-50-1461-fig007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8023/7491299/42d9175f4aa8/turkjmedsci-50-1461-fig008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8023/7491299/a50fccf59297/turkjmedsci-50-1461-fig009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8023/7491299/f547e12a8fda/turkjmedsci-50-1461-fig010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8023/7491299/68a9f06c8266/turkjmedsci-50-1461-fig001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8023/7491299/c84a99e1fc87/turkjmedsci-50-1461-fig002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8023/7491299/ccbfd4f7035f/turkjmedsci-50-1461-fig003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8023/7491299/14346415dc6e/turkjmedsci-50-1461-fig004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8023/7491299/25482dbe6fbc/turkjmedsci-50-1461-fig005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8023/7491299/43a82218e497/turkjmedsci-50-1461-fig006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8023/7491299/a21be9ba08d9/turkjmedsci-50-1461-fig007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8023/7491299/42d9175f4aa8/turkjmedsci-50-1461-fig008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8023/7491299/a50fccf59297/turkjmedsci-50-1461-fig009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8023/7491299/f547e12a8fda/turkjmedsci-50-1461-fig010.jpg

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