Tyrosyl radical in galactose oxidase not strongly perturbed by cysteine cross-link

Several density functional methods are applied to calculate the hyperfine c oupling constants and spin population distributions of sulfur-substituted a nd unsubstituted tyrosyl radical. The cysteine-substituted tyrosyl radical is found at the active site of the radical enzyme of galactose oxidase, The : main conclusion is that the sulfur center only possesses it small amount of unpaired spin not sufficient to perturb the overall odd-alternant spin d istribution in the tyrosyl radical. The original assignment of two hyperfin e couplings, one from a tyrosine beta-proton and one from H-5, is clearly c onfirmed by calculated isotropic hyperfine coupling constants. The inclusio n of solvent effects does not alter any of the conclusions. (C) 1999 Elsevi er Science B.V. All rights reserved.