title: Probing the metal ion selectivity in methionine aminopeptidase via changes in the luminescence properties of the enzyme bound europium ion.
abstract: We report herein, for the first time, that Europium ion (Eu(3+)) binds to the "apo" form of Escherichia coli methionine aminopeptidase (EcMetAP), and such binding results in the activation of the enzyme as well as enhancement in the luminescence intensity of the metal ion. Due to competitive displacement of the enzyme-bound Eu(3+) by different metal ions, we could determine the binding affinities of both "activating" and "non-activating" metal ions for the enzyme via fluorescence spectroscopy. The experimental data revealed that among all metal ions, Fe(2+) exhibited the highest binding affinity for the enzyme, supporting the notion that it serves as the physiological metal ion for the enzyme. However, the enzyme-metal binding data did not adhere to the Irving-William series. On accounting for the binding affinity vis a vis the catalytic efficiency of the enzyme for different metal ions, it appears evident that that the "coordination states" and the relative softness" of metal ions are the major determinants in facilitating the EcMetAP catalyzed reaction.
authors: Department of Chemistry and Biochemistry, North Dakota State University, Fargo, ND 58108, USA.
authors: Nitesh
authors: N
authors: Sule
authors: Raushan K
authors: RK
authors: Singh
authors: Pinjing
authors: P
authors: Zhao
authors: D K
authors: DK
authors: Srivastava
mesh_terms: D000626:Aminopeptidases / Q000737:chemistry* / Q000378:metabolism; D001667:Binding, Competitive; D055162:Biocatalysis; D020134:Catalytic Domain; D004789:Enzyme Activation; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism; D005063:Europium / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D008670:Metals / Q000737:chemistry / Q000378:metabolism; D063208:Methionyl Aminopeptidases; D008958:Models, Molecular; D013050:Spectrometry, Fluorescence
chemical_list: D029968:Escherichia coli Proteins; D008670:Metals; D005063:Europium; D000626:Aminopeptidases; D063208:Methionyl Aminopeptidases