Structure-function analyses of the ATX1 metallochaperone

被引:125
作者
Portnoy, ME
Rosenzweig, AC
Rae, T
Huffman, DL
O'Halloran, TV
Culotta, VC
机构
[1] Johns Hopkins Univ, Sch Publ Hlth, Dept Environm Hlth Sci, Baltimore, MD 21205 USA
[2] Johns Hopkins Univ, Sch Publ Hlth, Dept Biochem, Baltimore, MD 21205 USA
[3] Northwestern Univ, Dept Biochem Mol Biol & Cell Biol, Evanston, IL 60208 USA
[4] Northwestern Univ, Dept Chem, Evanston, IL 60208 USA
关键词
D O I
10.1074/jbc.274.21.15041
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Saccharomyces cerevisiae Atx1p represents a member of the family of metallochaperone molecules that escort copper to distinct intracellular targets, Atx1p specifically delivers copper to the Ccc2p copper transporter in the Golgi. Additionally, when overproduced, Atx1p substitutes for superoxide dismutase 1 in preventing oxidative damage; however the mechanistic overlap between these functions is unresolved. The crystal structure of Atx1p has been solved recently, By examining a surface electrostatic potential distribution, multiple conserved lysines are revealed on one face of Atx1p. An additional conserved lysine (Lys(65)) lies in close proximity to the metal binding site. Through site-directed mutagenesis, residues in the metal binding region including Lys65 were found to be necessary for both copper delivery to Ccc2p and for Atx1p antioxidant activity. Copper trafficking to Ccc2p also relied on the lysine rich face of Atx1p. Surprisingly however, elimination of these lysines did not inhibit the antioxidant activity of Atx1p, We provide evidence that Atx1p does not suppress oxidative damage by a metallochaperone mechanism but may directly consume superoxide. Purified Cu-Atx1p reacts noncatalytically with superoxide anion in vitro. We conclude that the copper-trafficking and antioxidant functions of Atx1p arise from chemically and structurally distinct attributes of this metallochaperone.
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页码:15041 / 15045
页数:5
相关论文
共 37 条
  • [21] Linder M. C., 1991, BIOCH COPPER, V1-13
  • [22] LIU XF, 1992, J BIOL CHEM, V267, P18298
  • [23] PROTEIN FOLDING AND ASSOCIATION - INSIGHTS FROM THE INTERFACIAL AND THERMODYNAMIC PROPERTIES OF HYDROCARBONS
    NICHOLLS, A
    SHARP, KA
    HONIG, B
    [J]. PROTEINS-STRUCTURE FUNCTION AND BIOINFORMATICS, 1991, 11 (04) : 281 - 296
  • [24] Expression of copper trafficking genes in the mouse brain
    Nishihara, E
    Furuyama, T
    Yamashita, S
    Mori, N
    [J]. NEUROREPORT, 1998, 9 (14) : 3259 - 3263
  • [25] Metal ion chaperone function of the soluble Cu(I) receptor Atx1
    Pufahl, RA
    Singer, CP
    Peariso, KL
    Lin, SJ
    Schmidt, PJ
    Fahrni, CJ
    Culotta, VC
    PennerHahn, JE
    OHalloran, TV
    [J]. SCIENCE, 1997, 278 (5339) : 853 - 856
  • [26] Undetectable intracellular free copper: The requirement of a copper chaperone for superoxide dismutase
    Rae, TD
    Schmidt, PJ
    Pufahl, RA
    Culotta, VC
    O'Halloran, TV
    [J]. SCIENCE, 1999, 284 (5415) : 805 - 808
  • [27] ROSENZWEIG AC, 1999, IN PRESS STRUCTURE L
  • [28] Sherman F, 1978, METHODS YEAST GENETI
  • [29] SIKORSKI RS, 1989, GENETICS, V122, P19
  • [30] A permease-oxidase complex involved in high-affinity iron uptake in yeast
    Stearman, R
    Yuan, DS
    YamaguchiIwai, Y
    Klausner, RD
    Dancis, A
    [J]. SCIENCE, 1996, 271 (5255) : 1552 - 1557