Deciphering the energy landscape of the interaction uranyl-DCP with antibodies using dynamic force spectroscopy.
Previous studies on molecular recognition of uranyl-DCP (dicarboxy-phenanthroline chelator) compound by two distinct monoclonal antibodies (Mabs U04S and U08S) clearly showed the presence of a biphasic shape in Bell-Evans' plots and an accentuated difference in slopes at the high loading rates. To further explore the basis in the slope difference, we have performed complementary experiments using antibody PHE03S, raised against uranyl-DCP but, presenting a strong cross-reactivity toward the DCP chelator. This work allowed us to obtain a reallocation of the respective contributions of the metal ion itself and that of the chelator. Results led us to propose a 2D schematic model representing two energy barriers observed in the systems Mabs U04S- and U08S-[UO(2)-DCP] where the outer barrier characterizes the interaction between UO(2) and Mab whereas the inner barrier characterizes the interaction between DCP and Mab. Using dynamic force spectroscopy, it is thus possible to dissect molecular interactions during the unbinding between proteins and ligands.
Teulon
Jean-Marie
JM
Commissariat à l'Energie Atomique, Institute for Biotechnology and Environmental Biology, Marcoule, F-30207 Bagnols sur Cèze, France.
Parot
Pierre
P
Odorico
Michael
M
Pellequer
Jean-Luc
JL
D000911:Antibodies, Monoclonal / Q000737:chemistry*; D001665:Binding Sites; D002614:Chelating Agents / Q000737:chemistry*; D004735:Energy Transfer; D018625:Microscopy, Atomic Force / Q000379:methods*; D010618:Phenanthrolines / Q000737:chemistry*; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods; D017974:Uranium Compounds / Q000737:chemistry*
D000911:Antibodies, Monoclonal; D002614:Chelating Agents; D010618:Phenanthrolines; D017974:Uranium Compounds
Biochimie. 2006 Nov;88(11):1605-18
16996675
Biophys J. 2007 Nov 15;93(10):3583-90
17675348
Ultramicroscopy. 2007 Oct;107(10-11):887-94
17644254
Biophys J. 2002 Oct;83(4):2270-9
12324444
Biophys J. 1997 Apr;72(4):1541-55
9083660
Science. 1978 May 12;200(4342):618-27
347575
Bioconjug Chem. 2004 Sep-Oct;15(5):1125-36
15366969
Biophys J. 2007 Jul 15;93(2):645-54
17449661
Toxicol Sci. 2008 Jun;103(2):397-408
18375546
Biochimie. 2006 Nov;88(11):1631-8
16815621
Chem Res Toxicol. 2008 Feb;21(2):349-57
18154273
Proc Natl Acad Sci U S A. 2000 Aug 29;97(18):9972-7
10963664
J Mol Recognit. 2007 Nov-Dec;20(6):508-15
18080996
Langmuir. 2007 Feb 27;23(5):2668-73
17269804