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Sensitive and Label-Free Detection of Protein Secondary Structure by Amide III Spectral Signals using Surface-Enhanced Raman Spectroscopy
Kang-zhen Tian,Chang-chun Cao,Xin-ming Nie*,Wen Wang,Cai-qin Han*
Author NameAffiliationE-mail
Kang-zhen Tian Jiangsu Key Laboratory of Advanced Laser Materials and Devices, School of Physics and Electronic Engineering, Jiangsu Normal University, Xuzhou 221116, China  
Chang-chun Cao The 95979 Army of Chinese People's Liberation Army, Taian 271200, China  
Xin-ming Nie* Jiangsu Key Laboratory of Advanced Laser Materials and Devices, School of Physics and Electronic Engineering, Jiangsu Normal University, Xuzhou 221116, China nxinming@jsnu.edu.cn 
Wen Wang Jiangsu Key Laboratory of Advanced Laser Materials and Devices, School of Physics and Electronic Engineering, Jiangsu Normal University, Xuzhou 221116, China  
Cai-qin Han* Jiangsu Key Laboratory of Advanced Laser Materials and Devices, School of Physics and Electronic Engineering, Jiangsu Normal University, Xuzhou 221116, China hancq@jsnu.edu.cn 
Abstract:
Proteins and peptides perform a vital role in living systems, however it remains a challenge for accurate description of proteins at the molecular level. Despite that surface-enhanced Raman spectroscopy (SERS) can provide the intrinsic fingerprint information of samples with ultrahigh sensitivity, it suffers from the poor reproducibility and reliability. Herein, we demonstrate that the silver nanorod array fabricated by an oblique angle deposition method is a powerful substrate for SERS to probe the protein secondary structures without exogenous labels. With this method, the SERS signals of two typical proteins (lysozyme and cytochrome c) are successfully obtained. Additionally, by analyzing the spectral signals of the amide III of protein backbone, the influence of concentration on the folding status of proteins has been elucidated. With the concentration increasing, the components of α-helix and β-sheet structures of lysozyme increase while the secondary structures of cytochrome c almost keep constant. The SERS method in this work offers an effective optical marker to characterize the structures of proteins.
Key words:  Surface-enhanced Raman spectroscopy, Silver nanorod, Protein secondary structures
FundProject:
表面增强拉曼光谱对于蛋白质二级结构的酰胺III谱带表征
田康振,曹长春,聂新明*,王 稳,韩彩芹*
摘要:
本文利用斜角沉积法制备银纳米棒阵列基底用于蛋白质二级结构的检测,结合酰胺III谱带光谱分析,建立了一种基于表面增强拉曼光谱(SERS)蛋白质二级结构的表征方法. 用这种方法获得了两种典型蛋白质(溶菌酶和细胞色素C)的SERS信号. 通过分析蛋白质骨架酰胺III的光谱,研究了浓度对蛋白质折叠状态的影响. 结果表明在一定范围内随着浓度的增加,溶菌酶的α-螺旋结构和β-折叠结构成分增加,而细胞色素C的二级结构基本保持不变.
关键词:  表面增强拉曼光谱,银纳米棒阵列基底,蛋白质二级结构
DOI:10.1063/1674-0068/cjcp1811267
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