College of Pharmacy, Ewha W. University, Seoul 03760, Republic of Korea; School of Computational Sciences, Korea Institute for Advanced Study, Seoul 02455, Republic of Korea.
College of Pharmacy, Ewha W. University, Seoul 03760, Republic of Korea.
Int J Biol Macromol. 2021 Jan 1;166:221-228. doi: 10.1016/j.ijbiomac.2020.10.152. Epub 2020 Oct 23.
Human serum albumin (HSA) is the most abundant protein in human plasma and plays versatile biological role. HSA has been widely used to treat several diseases and develop biocompatible biomaterials for biomedical applications. However, pharmaceutical-grade HSA (p-HSA) showed the altered oxidative and ligand-binding properties compare to native HSA. To investigate the influences of the manufacturing process on the molecular state of HSA, we determined the first crystal structure of p-HSA using the commercial HSA solution without any defatting step and further purification and carried out mass spectrometry to identify bound ligands. The crystal structure of p-HSA revealed that medium- and long-chain fatty acids and tryptophan are bound to p-HSA and one free cysteine is oxidized to cysteine-sulfenic acid. The mass spectra of p-HSA also confirmed the existence of fatty acids and tryptophan in p-HSA. Our results enhance understanding of the molecular state of p-HSA and can be utilized to produce p-HSA solutions and HSA-based biomaterials that has a higher biorelevance.
人血清白蛋白（HSA）是人体血浆中含量最丰富的蛋白质，具有多种生物学功能。HSA 已被广泛用于治疗多种疾病，并开发用于生物医学应用的生物相容性生物材料。然而，与天然 HSA 相比，药用级 HSA（p-HSA）表现出氧化和配体结合性质的改变。为了研究制造工艺对 HSA 分子状态的影响，我们使用未经脱脂步骤和进一步纯化的商业 HSA 溶液确定了 p-HSA 的第一个晶体结构，并进行了质谱分析以鉴定结合的配体。p-HSA 的晶体结构表明，中链和长链脂肪酸以及色氨酸结合到 p-HSA 上，一个游离半胱氨酸被氧化为半胱氨酸亚磺酸。p-HSA 的质谱也证实了 p-HSA 中存在脂肪酸和色氨酸。我们的结果增强了对 p-HSA 分子状态的理解，并可用于生产具有更高生物相关性的 p-HSA 溶液和基于 HSA 的生物材料。
