CRUI: Quantification of Metal Transfer From Metallothionein to Apometalloproteins Using Voltammetry Combined On Line with HPLC/ICP-MS

CRUI：使用伏安法与 HPLC/ICP-MS 结合在线定量从金属硫蛋白到脱辅基金属蛋白的金属转移

• 批准号: 9978806
• 负责人: Andrew Mason
• 金额: $ 103.62万
• 依托单位: California State University-Long Beach
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-01 至 2006-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9978806&HistoricalAwards=false
• 关键词: CRUI; Quantification; Metal; Transfer; Metallothionein

Life has evolved in close association with metals and it is not surprising, therefore, that evolutionary processes have selectively incorporated specific metals to take advantage of the additional chemical attributes that these metals bring to biological molecules. Thus, the incorporation of Zn or Cu metal may confer additional steric possibilities, catalytic activity or redox properties to the acceptor molecule and these metalloproteins are known to control important biological processes as varied as gene expression, respiration and acid/base balance.One intracellular protein that has been advocated to be involved in all of these processes in metallothionein, although the mechanisms by which this protein controls cellular metal homeostasis is unclear. A novel technique (EC/HPLC/ICP-MS) involving directly coupling electrochemistry (EC), high performance liquid chromatography (HPLC) and inductively coupled plasma mass spectroscopy (ICP-MS) will be developed to conduct a series of experiments that have been collectively designed to determine (i) the relative importance of glutathione and redox conditions necessary to cause the donation and acceptance of essential metals between metallothionein and apoenzymes within a complex cytoplasmic milieu; (ii) whether glutathione disulfide or oxidizing conditions can lead to the release of Cd and other physiologically toxic metals from metallothionein causing subsequent non-specific binding to target macromolecules.

生命的进化与金属密切相关，因此，进化过程选择性地结合特定的金属以利用这些金属为生物分子带来的额外化学属性也就不足为奇了。 因此，Zn或Cu金属的掺入可以赋予受体分子额外的空间可能性、催化活性或氧化还原性质，并且已知这些金属蛋白控制重要的生物过程，如基因表达、呼吸和酸/碱平衡。尽管这种蛋白质控制细胞金属稳态的机制尚不清楚。 一种新技术（EC/HPLC/ICP-MS）涉及直接耦合电化学（EC），将开发高效液相色谱法（HPLC）和电感耦合等离子体质谱法（ICP-MS），以进行一系列实验，这些实验共同设计用于确定（i）谷胱甘肽和氧化还原条件的相对重要性，必须引起捐赠和接受之间的金属硫蛋白和脱辅基酶在一个复杂的细胞质环境;（ii）谷胱甘肽二硫化物或氧化条件下是否可以导致释放镉和其他生理毒性金属硫蛋白引起随后的非特异性结合目标大分子。