title: Temperature and anation studies of the type 2 site in Rhus vernicifera laccase.
abstract: Temperature-dependent structural changes involving the type 2 site in laccase are probed by EPR studies of a derivative of laccase in which the type 1 Cu has been replaced by Hg(II) [Morie-Bebel, M. M., Morris, M. C., Menzie, J. L., & McMillin, D. R. (1984) J. Am. Chem. Soc. 106, 3677-3678]. At the temperature extremes (123 and 299 K), single well-defined species are present, but at intermediate temperatures (between 213 and 253 K), the presence of multiple structures is indicated. For the first time, the room temperature EPR spectrum of the type 2 copper has been resolved. Azide binding and fluoride binding have also been studied as a function of temperature. The results suggest that each anion preferentially interacts with the type 3 site in fluid solution and that these adducts can be trapped by rapidly cooling the sample to 123 K. Annealing the adducts at 253 K permits rearrangement and binding at an equatorial position of the type 2 Cu. This pathway to anation at the type 2 site contrasts sharply with previous studies which required a large excess of anions, and it reveals important insight into the flexibility of the type 2/type 3 cluster in laccase.
authors.affiliation: Department of Chemistry, Purdue University, West Lafayette, Indiana 47907-3699.
authors.forename: J C
authors.initials: JC
authors.lastname: Severns
authors.forename: D R
authors.initials: DR
authors.lastname: McMillin
mesh_terms: D004578:Electron Spin Resonance Spectroscopy; D042845:Laccase; D010088:Oxidoreductases / Q000378:metabolism*; D010940:Plant Proteins / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D013696:Temperature; D014197:Trees
chemical_list: D010940:Plant Proteins; D010088:Oxidoreductases; D042845:Laccase