The Metal Corrdination of sCD39 During ATP Hydrolysis
View/ Open
Guidotti_MetalCoordination.pdf (907.5Kb)
Access Status
Full text of the requested work is not available in DASH at this time ("restricted access"). For more information on restricted deposits, see our FAQ.Published Version
https://doi.org/10.1186/1471-2091-2-9Metadata
Show full item recordCitation
Chen, Wei, and Guido Guidotti. 2001. The metal coordination of sCD39 during ATP hydrolysis. BMC Biochemistry 2: 9.Abstract
BackgroundThe hydrolysis of ATP and ADP by ecto-nucleoside triphosphate diphosphohydrolase 1 (CD39) requires divalent cations, like Ca2+ and Mg2+. In spite of considerable work, it is not clear whether divalent cations bind to the enzyme in the absence of nucleotide or only as nucleotide-Me+2 complex. Here we study the protein ligands for Me+2.
Results
When VO2+ was used as a substitute for Ca2+, the ATPase activity of soluble CD39 was 25% of that with Ca2+ as cofactor. Protein ligands of the VO2+-nucleotide complex bound to the catalytic site of soluble CD39 were characterized by electron paramagnetic resonance (EPR) spectroscopy. The EPR spectrum contained one species designated T with VO2+-AMPPNP as ligand. Two species D1 and D2 were observed when VO2+-AMPCP was bound to soluble CD39. The results suggest that species D1 and D2 represent the metal-ADP complexes at the catalytic site of soluble CD39 corresponding to the intermediate formed during ATP hydrolysis and the substrate for further hydrolysis, respectively.
Conclusions
VO2+ can functionally substitute for Ca2+ as a cofactor of sCD39, and it produces four different EPR features when bound in the presence of different nucleotides or in the absence of nucleotide. The metal coordination for each conformation corresponding to each EPR species is proposed, and the mechanism of sCD39 catalysis is discussed.
Other Sources
http://www.biomedcentral.com/1471-2091/2/9Citable link to this page
http://nrs.harvard.edu/urn-3:HUL.InstRepos:3228040
Collections
- FAS Scholarly Articles [18292]
Contact administrator regarding this item (to report mistakes or request changes)