Site-specific covalent modifications of human insulin by catechol estrogens: Reactivity and induced structural and functional changes

Ming Chun Ku, Chieh Ming Fang, Juei Tang Cheng, Huei Chen Liang, Tzu Fan Wang, Chih Hsing Wu, Chiao Chen Chen, Jung Hsiang Tai, Shu Hui Chen

研究成果: Article

8 引文 (Scopus)

摘要

Proteins, covalently modified by catechol estrogens (CEs), were identified recently from the blood serum of diabetic patients and referred to as estrogenized proteins. Estrogenization of circulating insulin may occur and affect its molecular functioning. Here, the chemical reactivity of CEs towards specific amino acid residues of proteins and the structural and functional changes induced by the estrogenization of insulin were studied using cyclic voltammetry, liquid chromatography-mass spectrometry, circular dichroism spectroscopy, molecular modeling, and bioassays. Our results indicate that CEs, namely, 2-and 4-hydroxyl estrogens, were thermodynamically and kinetically more reactive than the catechol moiety. Upon co-incubation, intact insulin formed a substantial number of adducts with one or multiple CEs via covalent conjugation at its Cys 7 in the A or B chain, as well as at His10 or Lys29 in the B chain. Such conjugation was coupled with the cleavage of inter-chain disulfide linkages. Estrogenization on these sites may block the receptor-binding pockets of insulin. Insulin signaling and glucose uptake levels were lower in MCF-7 cells treated with modified insulin than in cells treated with native insulin. Taken together, our findings demonstrate that insulin molecules are susceptible to active estrogenization, and that such modification may alter the action of insulin.

原文English
文章編號28804
期刊Scientific reports
6
DOIs
出版狀態Published - 2016 六月 29

指紋

Catechol Estrogens
Insulin
Proteins
MCF-7 Cells
Circular Dichroism
Liquid Chromatography
Disulfides
Biological Assay
Hydroxyl Radical
Mass Spectrometry
Spectrum Analysis
Estrogens

All Science Journal Classification (ASJC) codes

  • General

引用此文

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abstract = "Proteins, covalently modified by catechol estrogens (CEs), were identified recently from the blood serum of diabetic patients and referred to as estrogenized proteins. Estrogenization of circulating insulin may occur and affect its molecular functioning. Here, the chemical reactivity of CEs towards specific amino acid residues of proteins and the structural and functional changes induced by the estrogenization of insulin were studied using cyclic voltammetry, liquid chromatography-mass spectrometry, circular dichroism spectroscopy, molecular modeling, and bioassays. Our results indicate that CEs, namely, 2-and 4-hydroxyl estrogens, were thermodynamically and kinetically more reactive than the catechol moiety. Upon co-incubation, intact insulin formed a substantial number of adducts with one or multiple CEs via covalent conjugation at its Cys 7 in the A or B chain, as well as at His10 or Lys29 in the B chain. Such conjugation was coupled with the cleavage of inter-chain disulfide linkages. Estrogenization on these sites may block the receptor-binding pockets of insulin. Insulin signaling and glucose uptake levels were lower in MCF-7 cells treated with modified insulin than in cells treated with native insulin. Taken together, our findings demonstrate that insulin molecules are susceptible to active estrogenization, and that such modification may alter the action of insulin.",
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Site-specific covalent modifications of human insulin by catechol estrogens : Reactivity and induced structural and functional changes. / Ku, Ming Chun; Fang, Chieh Ming; Cheng, Juei Tang; Liang, Huei Chen; Wang, Tzu Fan; Wu, Chih Hsing; Chen, Chiao Chen; Tai, Jung Hsiang; Chen, Shu Hui.

於: Scientific reports, 卷 6, 28804, 29.06.2016.

研究成果: Article

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AU - Fang, Chieh Ming

AU - Cheng, Juei Tang

AU - Liang, Huei Chen

AU - Wang, Tzu Fan

AU - Wu, Chih Hsing

AU - Chen, Chiao Chen

AU - Tai, Jung Hsiang

AU - Chen, Shu Hui

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