Active Transport of Manganese in Isolated Membranes of Escherichia coli

Accumulation of manganese was measured in subcellular membrane vesicles isolated from Escherichia coli. Accumulation of (54)Mn by vesicles in 0.5 m sucrose is stimulated by glucose and d-lactate and is inhibited by metabolic poisons such as dinitrophenol, m-chlorophenyl carbonylcyanide hydrazone, valinomycin, and nigericin. Manganese uptake by vesicles requires 10 mm calcium, which is not required for uptake of manganese by intact cells. The calcium requirement is specific and cannot be replaced by magnesium, sodium, or potassium. Strontium can replace calcium but is somewhat less effective than calcium. The uptake of manganese is via a manganese-specific system which shows saturation kinetics with manganese with a K(m) of 8 x 10(-6)m and a V(max) of 4 nmoles per min per g (wet weight) at 25 C. Magnesium and calcium do not compete for uptake. The accumulated manganese can be released from the vesicles by lipid active agents such as toluene, and can be exchanged for external manganese.