1UCN: X-Ray Structure Of Human Nucleoside Diphosphate Kinase A Complexed With Adp At 2 A Resolution

Citation:
Abstract
NDPK-A, product of the nm23-H1 gene, is one of the two major isoforms of human nucleoside diphosphate kinase. We analyzed the binding of its nucleotide substrates by fluorometric methods. The binding of nucleoside triphosphate (NTP) substrates was detected by following changes of the intrinsic fluorescence of the H118G/F60W variant, a mutant protein engineered for that purpose. Nucleoside diphosphate (NDP) substrate binding was measured by competition with a fluorescent derivative of ADP, following the fluorescence anisotropy of the derivative. We also determined an X-ray structure at 2.0A resolution of the variant NDPK-A in complex with ADP, Ca(2+) and inorganic phosphate, products of ATP hydrolysis. We compared the conformation of the bound nucleotide seen in this complex and the interactions it makes with the protein, with those of the nucleotide substrates, substrate analogues or inhibitors present in other NDP kinase structures. We also compared NDP kinase-bound nucleotides to ATP bound to protein kinases, and showed that the nucleoside monophosphate moieties have nearly identical conformations in spite of the very different protein environments. However, the beta and gamma-phosphate groups are differently positioned and oriented in the two types of kinases, and they bind metal ions with opposite chiralities. Thus, it should be possible to design nucleotide analogues that are good substrates of one type of kinase, and poor substrates or inhibitors of the other kind.
PDB ID: 1UCNDownload
MMDB ID: 24889
PDB Deposition Date: 2003/4/16
Updated in MMDB: 2012/10
Experimental Method:
x-ray diffraction
Resolution: 2  Å
Source Organism:
Similar Structures:
Biological Unit for 1UCN: hexameric; determined by author and by software (PISA,PQS)
Molecular Components in 1UCN
Label Count Molecule
Proteins (6 molecules)
6
Nucleoside Diphosphate Kinase a(Gene symbol: NME1)
Molecule annotation
Chemicals (24 molecules)
1
6
2
10
3
6
4
2
* Click molecule labels to explore molecular sequence information.

Citing MMDB
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