The regulation of catalytic activity of the menkes copper-translocating P-type ATPase - Role of high affinity copper-binding sites

被引:102
作者
Voskoboinik, I
Mar, J
Strausak, D
Camakaris, J [1 ]
机构
[1] Univ Melbourne, Dept Genet, Parkville, Vic 3010, Australia
[2] Deakin Univ, Sch Biol & Chem Sci, Ctr Cellular & Mol Biol, Burwood 3125, Australia
关键词
D O I
10.1074/jbc.M103532200
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
The Menkes protein is a transmembrane copper translocating P-type ATPase. Mutations in the Menkes gene that affect the function of the Menkes protein may cause Menkes disease in humans, which is associated with severe systemic copper deficiency. The catalytic mechanism of the Menkes protein, including the formation of transient acylphosphate, is poorly understood. We transfected and overexpressed wild-type and targeted mutant Menkes protein in yeast and investigated its transient acyl phosphorylation. We demonstrated that the Menkes protein is transiently phosphorylated by ATP in a copper-specific and copper-dependent manner and appears to undergo conformational changes in accordance with the classical P-type ATPase model. Our data suggest that the catalytic cycle of the Menkes protein begins with the binding of copper to high affinity binding sites in the transmembrane channel, followed by ATP binding and transient phosphorylation. We propose that. putative copper-binding sites at the N-terminal domain of the Menkes protein are important as sensors of low concentrations of copper but are not essential for the overall catalytic activity.
引用
收藏
页码:28620 / 28627
页数:8
相关论文
共 46 条
[1]  
Ausubel F.M., 1992, SHORT PROTOCOLS MOL, V2nd
[2]   Evolution of substrate specificities in the P-type ATPase superfamily [J].
Axelsen, KB ;
Palmgren, MG .
JOURNAL OF MOLECULAR EVOLUTION, 1998, 46 (01) :84-101
[3]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[4]   Molecular mechanisms of copper homeostasis [J].
Camakaris, J ;
Voskoboinik, I ;
Mercer, JF .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1999, 261 (02) :225-232
[5]   GENE AMPLIFICATION OF THE MENKES (MNK, ATP7A) P-TYPE ATPASE GENE OF CHO CELLS IS ASSOCIATED WITH COPPER RESISTANCE AND ENHANCED COPPER EFFLUX [J].
CAMAKARIS, J ;
PETRIS, MJ ;
BAILEY, L ;
SHEN, PY ;
LOCKHART, P ;
GLOVER, TW ;
BARCROFT, CL ;
PATTON, J ;
MERCER, JFB .
HUMAN MOLECULAR GENETICS, 1995, 4 (11) :2117-2123
[6]   CALCIUM-PUMP OF THE PLASMA-MEMBRANE [J].
CARAFOLI, E .
PHYSIOLOGICAL REVIEWS, 1991, 71 (01) :129-153
[7]   ISOLATION OF A CANDIDATE GENE FOR MENKES DISEASE THAT ENCODES A POTENTIAL HEAVY-METAL BINDING-PROTEIN [J].
CHELLY, J ;
TUMER, Z ;
TONNESEN, T ;
PETTERSON, A ;
ISHIKAWABRUSH, Y ;
TOMMERUP, N ;
HORN, N ;
MONACO, AP .
NATURE GENETICS, 1993, 3 (01) :14-19
[8]   Structure/function analysis of the amino-terminal region of the alpha 1 and alpha 2 subunits of Na,K-ATPase [J].
Daly, SE ;
Lane, LK ;
Blostein, R .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1996, 271 (39) :23683-23689
[9]  
Danks D.M., 1995, METABOLIC MOL BASIS, P2211
[10]   Expression, purification, and metal binding properties of the N-terminal domain from the Wilson disease putative copper-transporting ATPase (ATP7B) [J].
DiDonato, M ;
Narindrasorasak, S ;
Forbes, JR ;
Cox, DW ;
Sarkar, B .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1997, 272 (52) :33279-33282