| Autor: |
Koebke KJ; Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109, United States., Alfaro VS; Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109, United States., Pinter TBJ; Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109, United States., Deb A; Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109, United States., Lehnert N; Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109, United States., Tard C; LCM, CNRS, Ecole Polytechnique, IP Paris, F-91128 Palaiseau, France., Penner-Hahn JE; Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109, United States., Pecoraro VL; Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109, United States. |
| Abstrakt: |
Blue copper proteins have a constrained Cu(II) geometry that has proven difficult to recapitulate outside native cupredoxin folds. Previous work has successfully designed green copper proteins which could be tuned blue using exogenous ligands, but the question of how one can create a self-contained blue copper site within a de novo scaffold, especially one removed from a cupredoxin fold, remained. We have recently reported a red copper protein site within a three helical bundle scaffold which we later revisited and determined to be a nitrosocyanin mimic, with a CuHis 2 CysGlu binding site. We now report efforts to rationally design this construct toward either green or blue copper chromophores using mutation strategies that have proven successful in native cupredoxins. By rotating the metal binding site, we created a de novo green copper protein. This in turn was converted to a blue copper protein by removing an axial methionine. Following this rational sequence, we have successfully created red, green, and blue copper proteins within an alpha helical fold, enabling comparisons for the first time of their structure and function disconnected from the overall cupredoxin fold. |
