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Self-Assembling Protein Hydrogels with Modular Integrin Binding Domains

机译：具有模块整合素结合域的自组装蛋白水凝胶。

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Hydrogels with integrin binding activity were created from associating proteins with embedded RGD sequences.These proteins are a modified AC_(10)Bcys triblock design composed of acidic A and basic B leucine zipper associating domains flanking a new soluble disordered coil block that contains nine repeats of AGAGAGPEG and three copies of the RGD integrin binding sequence.As with the original AC_(10)Bcys design without the embedded RGD sequences,these proteins self-assemble into stable hydrogels at concentrations above approximately 50 mg/mL in a range of solution pH and temperature conditions.The mechanism for hydrogel assembly is the intermolecular association of A and B helical domains into bundles which act as cross-links connected by the soluble central disordered coil domains.The secondary structure of the proteins and the mechanical properties of the hydrogels they form are not adversely affected by the presence of the RGD sequences.The RGD sequences embedded in the disordered coil region support the adhesion,spreading,and polarization of human fibroblast cells on protein coated surfaces.Confocal microscopy studies demonstrated the presence of focal adhesion complexes and organized actin stress fibers in these cells.In contrast,fibroblasts seeded onto surfaces coated with the original AC_(10)Bcys protein remained rounded and did not form focal adhesions,indicating that bioactivity is conferred by the presence of the embedded RGD sequences.Such hydrogel-forming bioactive proteins have potential for cell and tissue culture applications.
机译：具有整合素结合活性的水凝胶是通过将蛋白质与嵌入的RGD序列关联而创建的，这些蛋白质是修饰的AC_（10）Bcys三嵌段设计，由酸性A和碱性B亮氨酸拉链缔合域组成，侧翼是一个新的可溶性无序卷曲嵌段，包含9个重复序列。 AGAGAGPEG和RGD整联蛋白结合序列的三个副本。与最初的AC_（10）Bcys设计没有嵌入的RGD序列一样，这些蛋白质在溶液pH和pH值范围内以高于约50 mg / mL的浓度自组装成稳定的水凝胶。水凝胶组装的机理是A和B螺旋结构域分子间缔合成束，这些束起可溶的中央无序卷曲结构域连接的交联作用。蛋白质的二级结构及其形成的水凝胶的机械性质不受RGD序列存在的不利影响。无序coi中嵌入的RGD序列l区支持人类成纤维细胞在蛋白涂层表面上的粘附，扩散和极化。共聚焦显微镜研究表明这些细胞中存在粘着斑复合物和组织化的肌动蛋白应力纤维。相反，成纤维细胞接种在涂有原始AC_的表面上（10）Bcys蛋白保持圆形，没有形成粘着斑，表明其生物活性是由嵌入的RGD序列赋予的。这种形成水凝胶的生物活性蛋白具有用于细胞和组织培养的潜力。

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