The Effect of Disulfide Bonds on Protein Folding, Unfolding, and Misfolding Investigated by FT-Raman Spectroscopy
學年 105
學期 1
出版（發表）日期 2016-08-06
作品名稱 The Effect of Disulfide Bonds on Protein Folding, Unfolding, and Misfolding Investigated by FT-Raman Spectroscopy
作品名稱（其他語言）
著者 Chih-Hsien Wang; Chia-Chi Huang; Long-Liu Lin; Wenlung Chen
單位
出版者
著錄名稱、卷期、頁數 Journal of Raman Spectroscopy 47(8), p.940-947
摘要 Disulfide bond is relevant to many protein folding/unfolding functions and conformational diseases. To elucidate the effects of disulfide bonds on protein folding, unfolding, and misfolding, we performed Fourier transform–Raman measurements on serial chemical‐induced denaturations of bovine serum albumin (BSA). By directly monitoring Raman stretching at S–S (~507 cm−1), S–H (~2566 cm−1), amide I (1655 cm−1 for α‐helix; 1667 cm−1 for β‐sheet structure), and amide III (>1300 cm−1 for α‐helix; 1246 cm−1 for β‐sheet structure), the status of disulfide bonds and secondary structure of BSA at different states were elucidated. Both disulfide bonds and secondary structure (mostly in α‐helix) of BSA appeared relatively stable even when the protein was unfolded by urea solution. However, disulfide bonds were completely reduced and protein secondary structure changed from α‐helix to a relatively β‐sheet dominant when the protein was modified by the mixed solution of urea and dithiothreitol (urea/DTT). Adhering to these structural changes, the protein proceeded to different degrees of polymerization. BSA would aggregate into a high molecular mass (over 700 kDa) of protein ensemble when it was exposed to the mixed urea/DTT solution. An irreversible change in S–S/S–H conversion and secondary structure was responsible for protein misfolding. We demonstrate here that Fourier transform–Raman directly probe S–S/S–H conversion and secondary structural change of BSA at different states, and these results clearly indicate that disulfide bonds and secondary structure of BSA serve as concrete frameworks to stabilize protein structure. As the frameworks collapse, the protein undergoes an irreversible structural change and results in protein misfolding.
關鍵字 disulfide bonds;FT–Raman;BSA;secondary structure;folding/unfolding
語言 en_US
ISSN 1097-4555
期刊性質 國外
收錄於 SCI
產學合作
通訊作者
審稿制度 是
國別 GBR
公開徵稿
出版型式 ,電子版,紙本
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