Four polyammonium macrocycles were synthesized and characterized: two with 21-membered rings and differing numbers of oxygen and nitrogen heteroatoms, [21]N6O (1) and [21]N5O2 (2), and two with bipyridine incorporated into the ring, [24]N4O2bipy (3) and [27]N3O2bipy (4). Their ability to catalyze the dephosphorylation of adenosine triphosphate was examined. It was found that ring size plays a crucial role in the catalytic ability of the macrocycles, with the 21-membered rings being superior to larger macrocycles. Also, rates of dephosphorylation were found to increase with increasing number of nitrogen atoms in the ring. For two of the macrocycles, crystal structures were determined. Macrocycle 2 crystallizes in the tricliriic space group PI, a = 10.692(1), b = 17.037(2), c = 8.1952(8)Å, a = 92.550(9), β = 100.816(9), γ = 106.77(1)°, V = 1396.1(3) Å3; the structure was solved to R = 0.089 and Rw = 0.098. Macrocycle 4 crystallizes in the monoclinic space group P21/n, a = 14.589(1), b = 15.427(1), c = 16.382(1) Å, b = 90.137(6)°, V = 3687.0(9) Å3; the structure was solved to R = 0.056 and Rw = 0.085.