The N-terminal domain of 2',3'-cyclic nucleotide 3'-phosphodiesterase harbors a GTP/ATP binding site.
The interaction between 2',3'-cyclic nucleotide 3'-phosphodiesterase and guanine/adenine nucleotides was investigated. The binding of purine nucleotides to 2',3'-cyclic nucleotide 3'-phosphodiesterase was revealed by both direct and indirect methods. In fact, surface plasmon resonance experiments, triphosphatase activity measurements, and fluorescence experiments revealed that 2',3'-cyclic nucleotide 3'-phosphodiesterase binds purine nucleotide triphosphates with an affinity higher than that displayed for diphosphates; on the contrary, the affinity for both purine monophosphates and pyrimidine nucleotides was negligible. An interpretation of biological experimental data was achieved by a building of 2',3'-cyclic nucleotide 3'-phosphodiesterase N-terminal molecular model. The structural elements responsible for nucleotide binding were identified and potential complexes between the N-terminal domain of CNP-ase and nucleotide were analyzed by docking simulations. Therefore, our findings suggest new functional and structural property of the N-terminal domain of CNPase.
Stingo
Stefania
S
Dipartimento di Scienze Farmaceutiche, Università di Salerno, Via Ponte Don Melillo, Fisciano (SA) 84084, Italy.
Masullo
Mariorosario
M
Polverini
Eugenia
E
Laezza
Chiara
C
Ruggiero
Immacolata
I
Arcone
Rosaria
R
Ruozi
Enrica
E
Dal Piaz
Fabrizio
F
Malfitano
Anna Maria
AM
D'Ursi
Anna Maria
AM
Bifulco
Maurizio
M
D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D001665:Binding Sites / Q000502:physiology; D003198:Computer Simulation; D008958:Models, Molecular; D011485:Protein Binding / Q000502:physiology; D011685:Purine Nucleotides / Q000737:chemistry* / Q000378:metabolism; D051381:Rats; D013329:Structure-Activity Relationship; D013379:Substrate Specificity / Q000502:physiology
D011685:Purine Nucleotides; D015087:2',3'-Cyclic-Nucleotide Phosphodiesterases