H53D+FAD complex H53D+FAD+dUMP complex Protein B-factors FAD B-factors Protein B-factors FAD B-factors dUMP B-factors Chain A 44.6 FAD-A 66.4 Chain A 31.6 FAD-A 65.0 UMP-A 23.9 Chain B 46.2 FAD-B 74.7 Chain B 34.7 FAD-B 64.2 UMP-B 26.9 Chain C 42.6 FAD-C 83.3 Chain C 29.4 Only one SO4 43.8 UMP-C 40.2 Chain D 43.8 FAD-D 85.8 Chain D 30.4 Only one SO4 48.2 Only PO4 57.6 Native+FAD complex (PDB code: 1O2A) Native+FAD+dUMP complex (PDB code: 1O26) Protein B-factors FAD B-factors Protein B-factors FAD B-factors dUMP B-factors Chain A 28.9 FAD-A 40.7 Chain A 32.3 FAD-A 22.2 UMP-A 22.2 Chain B 30.0 FAD -B 55.1 Chain B 33.6 FAD-B 20.0 UMP-B 20.2 Chain C 27.9 FAD-C 55.0 Chain C 31.1 FAD-C 22.6 UMP-C 23.0 Chain D 31.4 FAD-D 53.1 Chain D 32.3 FAD-D 20.7 UMP-D 21.7 aThe temperature factors generally show the positional movement of the atoms in the crystal structure. The molecules that are tightly bound in the protein show lower temperature factors compared to the molecules that interact weakly with the protein.

Table 2: Comparison of the temperature factors of various protein chains, FAD molecules and dUMP molecules in the mutant and native enzymea.