title: The calcium-binding site in the galactose chemoreceptor protein. Crystallographic and metal-binding studies.
abstract: We have determined the relative affinities in solution for various metals which bind to the lone calcium-binding site of the D-galactose-binding protein which resembles the EF-hand loop. In order of affinity the metals are: Ca2+ approximately Tb3+ approximately Pb2+ greater than Cd2+ greater than Sr2+ greater than Mg2+ much greater than Ba2+. The binding affinity for calcium (Kd = 2 microM) and the slow off-rate determined for terbium (1 x 10(-3) s-1) and that the metal-binding site is unperturbed by sugar binding argue for a structural role. Furthermore, we have crystallographically refined the structure of the binding protein with the calcium substituted by cadmium, compared it with the calcium-bound structure, and found them to be identical. The results of these structural and solution studies support the hypothesis that for a given metal-binding loop, cation hydration energy, size, and charge are major factors contributing to binding affinity.
authors.affiliation: Howard Hughes Medical Institute, Baylor College of Medicine, Houston, Texas 77030.
authors.forename: M N
authors.initials: MN
authors.lastname: Vyas
authors.forename: B L
authors.initials: BL
authors.lastname: Jacobson
authors.forename: F A
authors.initials: FA
authors.lastname: Quiocho
mesh_terms: D001665:Binding Sites; D002118:Calcium / Q000378:metabolism*; D002135:Calcium-Binding Proteins; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002413:Cations, Divalent; D004926:Escherichia coli / Q000378:metabolism; D005690:Galactose / Q000378:metabolism; D007700:Kinetics; D008958:Models, Molecular; D009004:Monosaccharide Transport Proteins; D033902:Periplasmic Binding Proteins; D011485:Protein Binding; D011487:Protein Conformation; D013725:Terbium / Q000378:metabolism
chemical_list: D002135:Calcium-Binding Proteins; D002352:Carrier Proteins; D002413:Cations, Divalent; D009004:Monosaccharide Transport Proteins; D033902:Periplasmic Binding Proteins; C023550:galactose-binding protein; D013725:Terbium; D002118:Calcium; D005690:Galactose