Near-infrared analysis of protein secondary structure in aqueous solutions and freeze-dried solids
Near-infrared analysis of protein secondary structure in aqueous solutions and freeze-dried solids
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DOI：
10.1002/jps.20580
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发表时间：
2006-04-01
影响因子：
3.8
通讯作者：
Aoyagi, N
中科院分区：
文献类型：
--
作者：
Izutsu, KI;Fujimaki, Y;Aoyagi, N
近红外光谱（NIR）的各种蛋白质（牛血清白蛋白，溶菌酶，卵清蛋白，γ-球蛋白，β-乳球蛋白，肌红蛋白，细胞色素c）的研究作为一种可能的分析方法的蛋白质二级结构在各种物理状态。蛋白质在水溶液中的光谱（透射模式，溶剂补偿）和那些在冷冻干燥固体（非破坏性漫反射模式）在组合（4000-5000 cm（-1））和第一泛音（5600-6600 cm（-1））光谱区域中在相似的频率显示几个带。水溶液中蛋白质的归一化二阶导数近红外光谱表明，一些谱带指示α-螺旋（4090、4365-4370、4615和5755 cm（-1））和β-折叠（4060、4405、4525-4540、4865和5915-5925 cm（-1））结构。蛋白质在冷冻干燥后大部分保持其天然结构的光谱特征，尽管观察到α-螺旋结构的一些减少和无序或β-折叠结构的增加。近红外分析还显示热处理BSA在水溶液和随后的冷冻干燥固体中的β-折叠形成。因此，目前的结果表明，无损近红外分析可用于研究脱水引起的蛋白质二级结构变化。(C)2006 Wiley-Liss，Inc.和美国药学协会药物科学杂志95：781-789，2006。
Near-infrared spectroscopy (NIR) of various proteins (bovine serum albumin, lysozyme, ovalbumin, gamma-globulin, beta-lactoglobulin, myoglobin, cytochrome-c) was investigated as a possible analytical method of the protein secondary structure in various physical states. The spectra of proteins in aqueous solutions (transmission mode, solvent-compensated) and those in freeze-dried solids (nondestructive diffuse reflection mode) showed several bands at similar frequencies in the combination (4000-5000 cm(-1)) and first overtone (5600-6600 cm(-1)) spectral regions. The normalized second-derivative near-infrared spectra of proteins in aqueous solutions suggested that some bands indicated alpha-helix (4090, 4365-4370, 4615, and 5755 cm(-1)) and beta-sheet (4060, 4405, 4525-4540, 4865, and 5915-5925 cm(-1)) structures. The proteins mostly maintained spectra characteristic of their native structure after freeze-drying, although some reductions in alpha-helical structure and increase in unordered or beta-sheet structures were observed. The near-infrared analysis also showed beta-sheet formation of heat-treated BSA in aqueous solutions and in subsequently freeze-dried solids. The present results thus indicated that the nondestructive near-infrared analysis can be used for the investigation of dehydration-induced changes in protein secondary structures. (C) 2006 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 95:781-789, 2006.