Comparison of the DING protein from the archaeon Sulfolobus solfataricus with human phosphate-binding protein and Pseudomonas fluorescence DING counterparts
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DING proteins represent a new group of 40 kDa-related members, ubiquitous in living organisms. The family also include the DING protein from Sulfolobus solfataricus, functionally related to poly(ADP-ribose) polymerases. Here, the archaeal protein has been compared with the human Phosphate-Binding Protein and the Pseudomonas fluorescence DING enzyme, by enzyme assays and immune cross-reactivity. Surprisingly, as the Sulfolobus enzyme, the Human and Pseudomonas proteins display poly(ADP-ribose) polymerase activity, whereas a phosphatase activity was only present in Sulfolobus and human protein, despite the conserved phosphate-binding site residues in Pseudomonas DING. All proteins were positive to anti-DING antibodies and gave a comparable pattern of anti-poly(ADP-ribose) polymerase immunoreactivity with two bands, at around 40 kDa and roughly at the double of this molecular mass. The latter signal was present in all Sulfolobus enzyme preparations and proved not due to either a contaminant or a precursor protein, but likely being a dimeric form of the 40 kDa polypeptide. The common immunological and partly enzymatic behavior linking human, Pseudomonas and Sulfolobus DING proteins, makes the archaeal protein an important model system to investigate DING protein function and evolution within the cell.
KeywordsP-binding proteins PfluDING HPBP PARPSso DING protein Sulfolobus solfataricus
M.R.F.M. thanks Dr. Barbara Nicolaus and “Servizio Fermentazioni” of ICB (CNR, Pozzuoli, Italy), for providing Sulfolobus MT-4 cells. M.R.F.M. is grateful to Dr. Eric Chabriere for providing HPBP protein and anti-HPBP antibodies, and to Dr. Ken Scott for gift of PfluDING and anti-PfluDING antibodies, and for their helpful comments to the manuscript. G.M. and E.P. acknowledge the support of Ministry of Instruction, University and Research (MIUR, PON 01_01585), and Flagship CNR project “Interomics” (subproject “Proteoma”) to G.M. ARH3 was a kind gift of Dr. M. Di Girolamo (Mario Negri Institute, S. Maria Imbaro, Chieti, Italy). Prof. A. Chiarugi (University of Florence, Italy) kindly provided PARP inhibitor (PJ34, N-(6-oxo-5,6-dihydrophenantridin2yl)N,N-dimethylacetamide-HCl).
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Conflict of interest
The authors declare no conflict of interest.
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