Nanomedicine and Drug Delivery Laboratory, Department of Pharmaceutical Sciences, University of Colorado Anschutz Medical Campus, Aurora, CO, USA.
Department of Ophthalmology, University of Colorado Anschutz Medical Campus, Aurora, CO, USA.
J Control Release. 2022 Dec;352:411-421. doi: 10.1016/j.jconrel.2022.10.026. Epub 2022 Oct 31.
Crystallins, small heat shock chaperone proteins that prevent protein aggregation, are of potential value in treating protein aggregation disorders. However, their therapeutic use is limited by their low potency and poor intracellular delivery. One approach to facilitate the development of crystallins is to improve their activity, stability, and delivery. In this study, zinc addition to αB-crystallin-D3 (αB-D3) formed supramolecular nano- and micro- assemblies, induced dose-dependent changes in structure (beta-sheet to alpha-helix) and increased surface hydrophobicity and chemical stability. Further, crystallin assemblies exhibited a size-dependent chaperone activity, with the nano-assemblies being superior to micro-assemblies and 4.3-fold more effective than the native protein in preventing β-mercaptoethanol induced aggregation of insulin. Insulin rescued by crystallin assemblies retained the activity as evidenced by glucose uptake in 3T3-L1 cells. The most active nano-assemblies enhanced protein stability, in the presence of urea, by 1.6-fold, whereas intracellular delivery was enhanced by 3.0-fold. The αB-D3 crystallin nano-assemblies exhibit uniquely enhanced stability, activity, and delivery compared to the native protein.
晶状蛋白是一种小型热休克伴侣蛋白，可防止蛋白质聚集，在治疗蛋白质聚集紊乱方面具有潜在价值。然而，其治疗用途受到其低效力和细胞内递送能力差的限制。促进晶状蛋白发展的一种方法是提高其活性、稳定性和递送能力。在这项研究中，锌的加入使αB-晶体蛋白-D3（αB-D3）形成超分子纳米和微组装体，诱导结构（β-折叠到α-螺旋）的剂量依赖性变化，并增加表面疏水性和化学稳定性。此外，晶状蛋白组装体表现出尺寸依赖性的伴侣活性，纳米组装体比微组装体优越，在防止β-巯基乙醇诱导胰岛素聚集方面比天然蛋白有效 4.3 倍。晶状蛋白组装体拯救的胰岛素保留了活性，这可以通过 3T3-L1 细胞中的葡萄糖摄取来证明。最活跃的纳米组装体在存在脲的情况下使蛋白质稳定性提高了 1.6 倍，而细胞内递送能力提高了 3.0 倍。与天然蛋白相比，αB-D3 晶状蛋白纳米组装体表现出独特的增强稳定性、活性和递送能力。
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