Bioactive site-specifically modified proteins for 4D patterning of gel biomaterials
Bioactive site-specifically modified proteins for 4D patterning of gel biomaterials
复制标题
DOI：
10.1038/s41563-019-0367-7
复制
发表时间：
2019-09-01
期刊：
影响因子：
41.2
通讯作者：
DeForest, Cole A.
中科院分区：
文献类型：
--
作者：
Shadish, Jared A.;Benuska, Gabrielle M.;DeForest, Cole A.
蛋白质修饰的生物材料可用于三维调节细胞功能。然而，随着对复杂细胞生理的动态异质控制的不断寻求，允许可逆和用户定义的脆性蛋白质与材料的捆绑的策略仍然非常需要。在这里，我们介绍了一种模块化和强大的半合成方法，用位点特异性修饰的蛋白质可逆地图案装载细胞的水凝胶。利用多用途的排序酶介导的转肽酶，我们生成了一个具有生物正交反应处理的均匀、单一功能化蛋白质的多样化文库，用于生物材料修饰。我们展示了光可逆的固定化荧光蛋白，酶和生长因子的凝胶具有优异的时空分辨率，同时保留天然蛋白的生物活性。局部表皮生长因子的表现可以动态调节增殖，细胞内有丝分裂原激活的蛋白激酶信号传导和亚细胞分解受体内吞作用。我们的方法广泛地允许对多种蛋白质进行修饰和模式化，这为在四个维度上探测和指导高级细胞命运提供了新的途径。
Protein-modified biomaterials can be used to modulate cellular function in three dimensions. However, as the dynamic heterogeneous control over complex cell physiology continues to be sought, strategies that permit a reversible and user-defined tethering of fragile proteins to materials remain in great need. Here we introduce a modular and robust semisynthetic approach to reversibly pattern cell-laden hydrogels with site-specifically modified proteins. Exploiting a versatile sortase-mediated transpeptidation, we generate a diverse library of homogeneous, singly functionalized proteins with bioorthogonal reactive handles for biomaterial modification. We demonstrate the photoreversible immobilization of fluorescent proteins, enzymes and growth factors to gels with excellent spatiotemporal resolution while retaining native protein bioactivity. Localized epidermal growth factor presentation enables dynamic regulation over proliferation, intracellular mitogen-activated protein kinase signalling and subcellularly resolved receptor endocytosis. Our method broadly permits the modification and patterning of a wide range of proteins, which provides newfound avenues to probe and direct advanced cellular fates in four dimensions.