Impact of the Molecular Environment on Thiol-Ene Coupling For Biofunctionalization and Conjugation
Impact of the Molecular Environment on Thiol-Ene Coupling For Biofunctionalization and Conjugation
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DOI：
10.1021/acs.bioconjchem.6b00349
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发表时间：
2016-09-01
影响因子：
4.7
通讯作者：
Gautrot, Julien E.
中科院分区：
文献类型：
--
作者：
Colak, Burcu;Da Silva, Julio C. S.;Gautrot, Julien E.
硫醇 - 烯自由基偶联越来越多地用于生物材料的生物功能化以及能够实现细胞包封的3D水凝胶的形成。实际上，硫醇 - 烯化学具有一些有趣的特性，对于需要肽或蛋白质进行特定反应的平台特别有吸引力，尤其是在细胞培养或包封过程中的原位反应。尽管人们对此很感兴趣，但对于在生物相关条件下原位影响硫醇 - 烯化学的因素知之甚少。在此，我们探索了在缓冲条件下控制光引发的硫醇 - 烯与一系列烯烃和硫醇（包括肽）偶联的一些分子参数。利用1H - 核磁共振（H - 1 NMR）和高效液相色谱（HPLC）来量化偶联效率以及缓冲液pH值的影响，以及靠近待偶联的烯烃和硫醇的分子结构和局部微环境的影响。其中一些观察结果得到了分子动力学和量子力学计算的支持。我们工作的一个重要发现是硫醇的pKa（及其随分子结构变化而产生的变化）对偶联效率有显著影响。同样，发现带正电荷的氨基酸和芳香族氨基酸对硫醇 - 烯偶联也有一定影响。因此，我们的研究表明，为了在含有肽的生物材料中实现高生物功能化水平或促进基于肽的水凝胶的高效形成，应仔细选择分子设计。
Thiol-ene radical coupling is increasingly used for the biofunctionalization of biomaterials and the formation of 3D hydrogels enabling cell encapsulation. Indeed, thiol-ene chemistry presents interesting features that are particularly attractive for platforms requiring specific reactions of peptides or proteins, in particular, in situ, during cell culture or encapsulation. Despite such interest, little is known about the factors impacting thiol-ene chemistry in situ, under biologically relevant conditions. Here we explore some of the molecular parameters controlling photoinitiated thiol-ene couplings with a series of alkenes and thiols, including peptides, in buffered conditions. H-1 NMR and HPLC were used to quantify the efficiency of couplings and the impact of the pH of the buffer, as well as the molecular structure and local microenvironment close to alkenes and thiols to be coupled. Some of these observations are supported by molecular dynamics and quantum mechanics calculations. An important finding of our work is that the plc of thiols (and its variation upon changes in molecular structure) have a striking impact on coupling efficiencies. Similarly, positively charged and aromatic amino acids are found to have some impact on thiol-ene couplings. Hence, our study demonstrates that molecular design should be carefully selected in order to achieve high biofunctionalization levels in biomaterials with peptides or promote the efficient formation of peptide-based hydrogels.