title: A requirement for calcium in the transport of cobalamin across the outer membrane of Escherichia coli.
abstract: Cobalamin (Cbl) transport across the outer membrane of cells of Escherichia coli consists of high affinity Cbl binding to the btuB protein of the Cbl receptor, followed by the proton motive force- and tonB-dependent release of the Cbl into the periplasmic space. During a search for experimental conditions that would mimic this release in vitro with isolated cell envelope particles, we found that calcium was required for the high affinity Cbl binding, and subsaturating calcium concentrations resulted in the decreased affinity of the Cbl receptor for Cbl. The apparent affinity of the Cbl receptor for calcium (KD, approximately 30 nM at pH 6.6) decreased with decreasing pH, resulting in decreased affinity for Cbl at lower pH values. With suboptimal levels of calcium, Cbl binding was decreased by millimolar levels of magnesium.
mesh_terms: D001693:Biological Transport, Active; D002118:Calcium / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D014805:Vitamin B 12 / Q000378:metabolism*
chemical_list: D014805:Vitamin B 12; D002118:Calcium