Fluorescent probes for the structure and function of metallothionein
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- 作者：Wolfgang Maret
- 关键词：Thiols ; Chemical modification ; Fluorescence ; Metallothionein ; Zinc
- 刊名：Journal of Chromatography B, Analytical Technologies in the Biomedical and Life Sciences
- 出版年：2009
- 出版时间：15 October 2009
- 年：2009
- 卷：877
- 期：28
- 页码：3378-3383
- 全文大小：369 K
文摘
Fluorescence methods have been instrumental in demonstrating that the structure of human metallothionein in vivo depends on the availability of metal ions and the redox environment. Differential chemical modifications of its cysteine thiols with fluorescent probes allowed determination of three states: metallothionein (zinc-bound thiolate), thionein (free thiols), and thionin (disulfides). Interrogation of its zinc-binding properties with fluorescent chelating agents revealed that the affinities for the seven zinc ions vary over four orders of magnitude. Attachment of fluorescent labels generated metallothionein FRET (fluorescence resonance energy transfer) sensors for investigating its structure and function in living cells.