Ghosh Ranendu, Sharma Sunny, Chattopadhyay Krishnananda
Structural Biology and Bioinformatics, Indian Institute of Chemical Biology, Council of Scientific and Industrial Research, 4, Raja S. C. Mullick Road, Kolkata 700032, India.
Biochemistry. 2009 Feb 10;48(5):1135-43. doi: 10.1021/bi802065j.
Arginine has been used extensively as an excipient in the formulation development of protein-based biopharmaceuticals. We investigate the role of arginine in suppressing protein aggregation and its mechanism by using bovine serum albumin as a model system. By using sedimentation velocity and other analytical techniques, we show that the use of arginine inhibits temperature-induced aggregation of the protein. We use fluorescence correlation spectroscopy and other spectroscopic techniques to show that arginine inhibits accumulation of partially folded intermediates, potentially involved in the aggregation process. The hydrodynamic radii of the protein in its native, unfolded, and intermediate states have been determined using fluorescence correlation spectroscopy at single-molecule resolution. A possible mechanism of the effects of arginine and its role as an aggregation suppressor has been discussed.
精氨酸已被广泛用作基于蛋白质的生物制药制剂开发中的辅料。我们以牛血清白蛋白为模型系统，研究精氨酸在抑制蛋白质聚集方面的作用及其机制。通过沉降速度法和其他分析技术，我们表明精氨酸的使用可抑制蛋白质的温度诱导聚集。我们使用荧光相关光谱法和其他光谱技术表明，精氨酸可抑制可能参与聚集过程的部分折叠中间体的积累。已在单分子分辨率下使用荧光相关光谱法测定了蛋白质在其天然、未折叠和中间状态下的流体动力学半径。本文讨论了精氨酸作用的可能机制及其作为聚集抑制剂的作用。
