title: Synthesis of bisphosphonate derivatives of ATP by T4 DNA ligase, ubiquitin activating enzyme (E1) and other ligases.
abstract: T4 DNA ligase and the ubiquitin activating enzyme (E1), catalyze the synthesis of ATP beta,gamma-bisphosphonate derivatives. Concerning T4 DNA ligase: (i) etidronate (pC(OH)(CH(3))p) displaced the AMP moiety of the complex E-AMP in a concentration dependent manner; (ii) the K(m) values and the rate of synthesis k(cat) (s(-1)), determined for the following compounds were, respectively: etidronate, 0.73+/-0.09 mM and (70+/-10)x10(-3) s(-1); clodronate (pCCl(2)p), 0.08+/-0.01 mM and (4.1+/-0.3)x10(-3) s(-1); methylenebisphosphonate (pCH(2)p), 0.024+/-0.001 mM and (0.6+/-0.1)x10(-3) s(-1); tripolyphosphate (P(3)) (in the synthesis of adenosine 5'-tetraphosphate, p(4)A), 1.30+/-0.30 mM and (6.2+/-1.1)x10(-3) s(-1); (iii) in the presence of GTP and ATP, inhibition of the synthesis of Ap(4)G was observed with clodronate but not with pamidronate (pC(OH)(CH(2)-CH(2)-NH(3))p). Concerning the ubiquitin activating enzyme (E1): methylenebisphosphonate was the only bisphosphonate, out of the ones tested, that served as substrate for the synthesis of an ATP derivative (K(m)=0.36+/-0.09 mM and k(cat)=0.15+/-0.02 s(-1)). None of the above bisphosphonates were substrates of the reaction catalyzed by luciferase or by acyl-CoA synthetase. The ability of acetyl-CoA synthetase to use methylenebisphosphonate as substrate depended on the commercial source of the enzyme. In our view this report widens our knowledge of the enzymes able to metabolize bisphosphonates, a therapeutic tool widely used in the treatment of osteoporosis.
authors: Departamento de Bioquímica, Instituto de Investigaciones Biomédicas Alberto Sols UAM/CSIC, Facultad de Medicina, 28029 Madrid, Spain.
authors: María A
authors: MA
authors: Günther Sillero
authors: Anabel
authors: A
authors: de Diego
authors: Francisco J
authors: FJ
authors: Pérez-Zúñiga
authors: Antonio
authors: A
authors: Sillero
mesh_terms: D000106:Acetate-CoA Ligase / Q000737:chemistry / Q000378:metabolism; D000105:Acetyl Coenzyme A / Q000737:chemistry / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000737:chemistry* / Q000378:metabolism; D011088:DNA Ligases / Q000378:metabolism; D004164:Diphosphonates / Q000737:chemistry* / Q000378:metabolism; D008156:Luciferases / Q000737:chemistry / Q000378:metabolism; D044764:Ubiquitin-Activating Enzymes / Q000737:chemistry / Q000378:metabolism
chemical_list: D004164:Diphosphonates; D000105:Acetyl Coenzyme A; D000255:Adenosine Triphosphate; D008156:Luciferases; D000106:Acetate-CoA Ligase; D044764:Ubiquitin-Activating Enzymes; D011088:DNA Ligases