C-Terminal Domain of the Membrane Copper Transporter Ctr1 from Saccharomyces cerevisiae Binds Four Cu(I) Ions as a Cuprous-Thiolate Polynuclear Cluster: Sub-femtomolar Cu(I) Affinity of Three Proteins Involved in Copper Trafficking

化学 酿酒酵母 酵母 离解常数 结晶学 电喷雾电离 胞浆 离解(化学) 质谱法 立体化学 分析化学(期刊) 生物化学 色谱法 受体 物理化学 有机化学
作者
Zhiguang Xiao,Fionna E. Loughlin,Graham N. George,Geoffrey J. Howlett,Anthony G. Wedd
出处
期刊:Journal of the American Chemical Society [American Chemical Society]
卷期号:126 (10): 3081-3090 被引量:240
标识
DOI:10.1021/ja0390350
摘要

The cytosolic C-terminal domain of the membrane copper transporter Ctr1 from the yeast Saccharomyces cerevisiae, Ctr1c, was expressed in E. coli as an oxygen-sensitive soluble protein with no significant secondary structure. Visible-UV spectroscopy demonstrated that Ctr1c bound four Cu(I) ions, structurally identified as a Cu(I)(4)(micro-S-Cys)(6) cluster by Xray absorption spectroscopy. This was the only metalated form detected by electrospray ionization mass spectrometry. An average dissociation constant K(D) = (K(1)K(2)K(3)K(4))(1/4) = 10(-)(19) for binding of Cu(I) to Ctr1c was estimated via competition with the ligand bathocuproine disulfonate bcs (beta(2) = 10(19.8)). Equivalent experiments for the yeast chaperone Atx1 and an N-terminal domain of the yeast Golgi pump Ccc2, which both bind a single Cu(I) ion, provided similar K(D) values. The estimates of K(D) were supported by independent estimates of the equilibrium constants K(ex) for exchange of Cu(I) between pairs of these three proteins. It is apparent that, in vitro, the three proteins buffer "free" Cu(I) concentrations in a narrow range around 10(-)(19) M. The results provide quantitative support for the proposals that, in yeast, (a) "free" copper concentrations are very low in the cytosol and (b) the Cu(I) trafficking gradient is shallow along the putative Ctrlc --> Atx1 --> Ccc2n metabolic pathway. In addition, both Ctr1c and its copper-responsive transcription factor Mac1 contain similar clusters which may be important in signaling copper status in yeast.
最长约 10秒,即可获得该文献文件

科研通智能强力驱动
Strongly Powered by AbleSci AI
更新
PDF的下载单位、IP信息已删除 (2025-6-4)

科研通是完全免费的文献互助平台,具备全网最快的应助速度,最高的求助完成率。 对每一个文献求助,科研通都将尽心尽力,给求助人一个满意的交代。
实时播报
刚刚
3秒前
3秒前
茸茸茸发布了新的文献求助30
3秒前
4秒前
迷路的沛芹完成签到 ,获得积分10
4秒前
香蕉觅云应助zedhumble采纳,获得10
7秒前
大道无形我有型完成签到,获得积分10
7秒前
9秒前
茸茸茸完成签到,获得积分10
9秒前
sda发布了新的文献求助10
9秒前
10秒前
ZZZ完成签到,获得积分10
11秒前
大个应助sda采纳,获得10
11秒前
12秒前
13秒前
Owen应助咕噜咕噜咕嘟咕嘟采纳,获得10
13秒前
14秒前
量子星尘发布了新的文献求助10
14秒前
m7m完成签到,获得积分10
14秒前
uniondavid完成签到,获得积分10
14秒前
16秒前
17秒前
魔幻的笑珊完成签到,获得积分10
17秒前
乐乐应助trocars采纳,获得10
18秒前
脑洞疼应助闾丘山菡采纳,获得10
19秒前
江筱筱完成签到,获得积分10
20秒前
天真之桃完成签到,获得积分10
21秒前
23秒前
Dmooou完成签到,获得积分10
24秒前
24秒前
27秒前
Rondab应助勤恳的夏之采纳,获得10
28秒前
29秒前
trocars发布了新的文献求助10
29秒前
Amos完成签到,获得积分10
30秒前
Rondab应助WQ采纳,获得10
30秒前
坚定的芷珊完成签到,获得积分10
30秒前
zedhumble发布了新的文献求助10
32秒前
大罗发布了新的文献求助10
33秒前
高分求助中
A new approach to the extrapolation of accelerated life test data 1000
ACSM’s Guidelines for Exercise Testing and Prescription, 12th edition 500
‘Unruly’ Children: Historical Fieldnotes and Learning Morality in a Taiwan Village (New Departures in Anthropology) 400
Indomethacinのヒトにおける経皮吸収 400
Phylogenetic study of the order Polydesmida (Myriapoda: Diplopoda) 370
基于可调谐半导体激光吸收光谱技术泄漏气体检测系统的研究 350
Robot-supported joining of reinforcement textiles with one-sided sewing heads 320
热门求助领域 (近24小时)
化学 材料科学 医学 生物 工程类 有机化学 生物化学 物理 内科学 纳米技术 计算机科学 化学工程 复合材料 遗传学 基因 物理化学 催化作用 冶金 细胞生物学 免疫学
热门帖子
关注 科研通微信公众号,转发送积分 3988997
求助须知:如何正确求助?哪些是违规求助? 3531351
关于积分的说明 11253520
捐赠科研通 3269928
什么是DOI,文献DOI怎么找? 1804830
邀请新用户注册赠送积分活动 882063
科研通“疑难数据库(出版商)”最低求助积分说明 809068