How the imidazole ring modulates amyloid formation of islet amyloid polypeptide: A chemical modification study

• Published in: Biochimica et biophysica acta Vol. 1860; no. 4; pp. 719 - 726
• Main Authors: Zhang, Xin, Liu, Junjun, Huang, lianqi, Yang, Xin, Petersen, Robert B., Sun, Yue, Gong, Hao, Zheng, Ling, Huang, Kun
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.04.2016
• Subjects: Amyloid; circular dichroism spectroscopy; Diethyl Pyrocarbonate - chemistry; ethyl acetate; fluorescence; fluorescent dyes; Fourier transform infrared spectroscopy; hIAPP; histidine; Humans; hydrogen bonding; Imidazole ring; Imidazoles - chemistry; Islet Amyloid Polypeptide - chemistry; Islet Amyloid Polypeptide - genetics; Islet Amyloid Polypeptide - metabolism; Modification; molecular dynamics; Molecular Dynamics Simulation; Multiprotein Complexes - chemistry; noninsulin-dependent diabetes mellitus; polypeptides; Protein Structure, Secondary; Thiazoles - chemistry; transmission electron microscopy; Imidazole ring; DMI; ThT; Modification; HFIP; IAPP; Amyloid; hIAPP; DEPC
• ISSN: 0304-4165; 0006-3002; 1872-8006
• DOI: 10.1016/j.bbagen.2016.01.008

The misfolding of human islet amyloid polypeptide (hIAPP) is an important pathological factor on the onset of type 2 diabetes. A number of studies have been focused on His18, the only histidine of hIAPP, whose imidazole ring and the protonation state might impact hIAPP amyloid formation, but the exact mechanism remains unclear. We used diethylpyrocarbonate (DEPC) to specifically modify His18 and obtained mono-ethyloxyformylated hIAPP (DMI). Thioflavin T based fluorescence, transmission electronic microscopy, circular dichroism spectroscopy, fluorescence dye leakage, Fourier transform infrared spectroscopy and replica-exchange molecular dynamics (REMD) simulation were applied to study the impact of DEPC-modification on hIAPP amyloid formation. After an ethyl-acetate group was introduced to the His18 of hIAPP by diethylpyrocarbonate (DEPC) modification, the pH dependent hIAPP fibrillation went to the opposite order and the number of intra-molecular hydrogen bonds decreased, while the possibility of His18 participating in the formation of α-helical structures increased. Furthermore, the membrane–peptide interaction and ion–peptide interaction were both impaired. The intramolecular hydrogen bond formation by His18 and the possibility of His18 participating in the formation of α-helical structures greatly modulated the manner of hIAPP amyloid formation. The imidazole ring directly participates in the hIAPP–membrane/ion interaction. DEPC modification is an alternative approach to investigate the role of the imidazole ring during amyloid formation. •We use DEPC to specifically modify and obtain mono-ethyloxyformylated hIAPP (DMI).•After DEPC modification, the pH dependent hIAPP fibrillation went to opposite order.•DEPC modification decreases the amount intra-molecular hydrogen bonds formed.•DEPC modification impairs the membrane–peptide interaction.•DEPC modification impairs the ion–peptide interaction.