The Binding Mode of ATP Revealed by the Solution Structure of the N-domain of Human ATP7A

Banci, Lucia; Bertini, Ivano; Cantini, Francesca; Inagaki, Sayaka; Migliardi, Manuele; Rosato, Antonio

doi:10.1074/jbc.m109.054262

Cited by 24 publications

(28 citation statements)

References 43 publications

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“…ATP binding locks the protein in the closed conformation and so does crystallization under the conditions used for the CopA ATP binding domain. In the N-domain of ATP7A, which is highly homologous to ATP7B, ATP binding primarily affects the loop connecting the ␣1-␣2 helices and, to a lesser extent, the orientation of these helices (5). In contrast, in the structure of WND-E1064A⌬, the ␣1-␣2 hairpin is markedly displaced compared with its position in the wild-type N-domain.…”

Section: Discussionmentioning

confidence: 77%

“…This observation strongly suggests the conformational mobility of the hairpin. A conformational change was also detected in the ATP7A N-domain in response to ATP binding (5). In the E1064A mutant, the orientation of the ␣1-␣2 hairpin is further altered and is clearly outside the range of the variations between the wild-type apo domains.…”

Section: ␣1-␣2 Hairpin Is the Most Variable Structural Element Of Thementioning

confidence: 80%

“…In contrast, NMR analysis of the N-domains from human ATP7B and ATP7A suggested protein flexibility (4,5). In solution, ATP binding to the N-domain appears to produce a more compact and stable structure (4,5), an observation further supported by recent thermal denaturation and molecular dynamics simulation experiments (9).…”

mentioning

confidence: 76%

“…The bulk of the ATP molecule is bound to the N-domain (3,4), whereas the ␥-phosphate extends to the P-domain to transiently phosphorylate the invariant aspartate residue Asp 1027 . The structure of the N-domain has been solved for human ATP7B (4) and several orthologs (5)(6)(7)(8). These studies have identified invariant residues contributing to the ATP binding pocket.…”

mentioning

confidence: 99%

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Difference in Stability of the N-domain Underlies Distinct Intracellular Properties of the E1064A and H1069Q Mutants of Copper-transporting ATPase ATP7B

Dmitriev

Bhattacharjee

Nokhrin

et al. 2011

Journal of Biological Chemistry

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Wilson disease (WD) is a disorder of copper metabolism caused by mutations in the Cu-transporting ATPase ATP7B. WD is characterized by significant phenotypic variability, the molecular basis of which is poorly understood. The E1064A mutation in the N-domain of ATP7B was previously shown to disrupt ATP binding. We have now determined, by NMR, the structure of the N-domain containing this mutation and compared properties of E1064A and H1069Q, another mutant with impaired ATP binding. The E1064A mutation does not change the overall fold of the N-domain. However, the position of the ␣1,␣2-helical hairpin (␣-HH) that houses Glu 1064 and His 1069 is altered. The ␣-HH movement produces a more open structure compared with the wild-type ATP-bound form and misaligns ATP coordinating residues, thus explaining complete loss of ATP binding. In the cell, neither the stability nor targeting of ATP7B-E1064A to the trans-Golgi network differs significantly from the wild type. This is in a contrast to the H1069Q mutation within the same ␣-HH, which greatly destabilizes protein both in vitro and in cells. The difference between two mutants can be linked to a lower stability of the ␣-HH in the H1069Q variant at the physiological temperature. We conclude that the structural stability of the N-domain rather than the loss of ATP binding plays a defining role in the ability of ATP7B to reach the trans-Golgi network, thus contributing to phenotypic variability in WD.The human copper-transporting ATPase ATP7B regulates copper levels in the cytosol and within the secretory pathway (1). Genetic mutations that disrupt the transport function of ATP7B lead to Wilson disease (WD), 4 a severe metabolic disorder associated with copper accumulation in the liver, brain, and other tissues (2). The WD phenotypes are rather diverse. The disease is characterized by significant variability in the age of onset, severity, and manifestations, which include hepatic, neurological, and psychiatric symptoms. More than 350 WDcausing mutations have been described. The majority of these mutations are single base pair substitutions that do not eliminate synthesis of the full-length ATP7B. Despite considerable effort, no strong correlations have been observed between specific mutations and the disease phenotype, necessitating better understanding of the effects that mutations have on ATP7B structure, function, and intracellular behavior.ATP7B transports copper from the cytosol across cellular membranes using the energy of ATP hydrolysis. ATP binds to the cytosolic ATP-hydrolyzing domain, which consists of the N-and P-domains. The bulk of the ATP molecule is bound to the N-domain (3, 4), whereas the ␥-phosphate extends to the P-domain to transiently phosphorylate the invariant aspartate residue Asp 1027 . The structure of the N-domain has been solved for human ATP7B (4) and several orthologs (5-8). These studies have identified invariant residues contributing to the ATP binding pocket. In ATP7B, these residues are Glu 1064 , Glu 1068 , His 1069 , Gly 1101 , Asn 115...

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Section: Discussionmentioning

confidence: 77%

Section: ␣1-␣2 Hairpin Is the Most Variable Structural Element Of Thementioning

confidence: 80%

mentioning

confidence: 76%

mentioning

confidence: 99%

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Difference in Stability of the N-domain Underlies Distinct Intracellular Properties of the E1064A and H1069Q Mutants of Copper-transporting ATPase ATP7B

Dmitriev

Bhattacharjee

Nokhrin

et al. 2011

Journal of Biological Chemistry

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“…Even discrete mutations within the NMBD sequence were shown to interfere with targeting and Golgi retention of nascent ATP7B (24). In fact, the rather extended NMBD sequence is flexible and undergoes functionally relevant conformational interactions as shown by its structural resolution by NMR spectroscopy (25,26), its propensity to interact with chaperone proteins (27), and the influence of its copper site redox status on overall conformation (28). Finally, we found that mutation of the serine residues undergoing phosphorylation interfered strongly with formation of CTV, underscoring the role of PKD-sustained phosphorylation in the final trafficking step.…”

Section: Discussionmentioning

confidence: 99%

Involvement of Protein Kinase D in Expression and Trafficking of ATP7B (Copper ATPase)

Pilankatta

Lewis

Inesi

2011

Journal of Biological Chemistry

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Copper TransportingATPases in Mammalian Cells

Yang

Lutsenko

2004

Encyclopedia of Inorganic and Bioinorganic Chemistry

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Copper‐transporting ATP ases ( Cu‐ATPases ) are essential for growth and development of mammalian organisms. These ubiquitous and evolutionarily conserved transporters participate in the biosynthesis of copper‐dependent enzymes and maintain copper levels within the range necessary to meet metabolic demands and avoid toxicity. Cu‐ATPases are the key components of sophisticated cellular machinery that evolved to provide precise inter‐ and intracellular distribution of metals, and the function of these transporters is tightly regulated. Copper along with a growing number of cytosolic proteins controls the activity, stability, and localization of Cu‐ATPases ; this regulation ensures balance between the delivery of copper to metalloenzymes in various cell compartments and copper export. Inactivating mutations in human Cu‐ ATPases ATP7A and ATP7B are associated with debilitating and often lethal disorders (Menkes disease and Wilson's disease, respectively). Biochemical and biophysical studies of ATP7A and ATP7B revealed multiple metal‐binding sites, complex multi‐domain architecture, and a significant role of interdomain interactions in the function and regulation of these transporters. This article summarizes the current data on the structure and mechanism of human Cu‐ ATPases ATP7A and ATP7B , as well as the biochemical basis of their regulation.

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The Binding Mode of ATP Revealed by the Solution Structure of the N-domain of Human ATP7A

Cited by 24 publications

References 43 publications

Difference in Stability of the N-domain Underlies Distinct Intracellular Properties of the E1064A and H1069Q Mutants of Copper-transporting ATPase ATP7B

Difference in Stability of the N-domain Underlies Distinct Intracellular Properties of the E1064A and H1069Q Mutants of Copper-transporting ATPase ATP7B

Involvement of Protein Kinase D in Expression and Trafficking of ATP7B (Copper ATPase)

Copper TransportingATPases in Mammalian Cells

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