Azurin as a Protein Scaffold for a Low-coordinate Nonheme Iron Site with a Small-molecule Binding Pocket

作者：McLaughlin Matthew P; Retegan Marius; Bill Eckhard; Payne Thomas M; Shafaat Hannah S; Pena Salvador; Sudhamsu Jawahar; Ensign Amy A; Crane Brian R; Neese Frank; Holland Patrick L^*

来源：Journal of the American Chemical Society, 2012, 134(48): 19746-19757.

DOI：10.1021/ja308346b

The apoprotein of Pseudomonas aeruginosa azurin binds iron(II) to give a 1:1 complex, which has been characterized by electronic absorption, Mossbauer, and NMR spectroscopies, as well as X-ray crystallography and quantum-chemical computations. Despite potential competition by water and other coordinating residues, iron(II) binds tightly to the low-coordinate site. The iron(II) complex does not react with chemical redox agents to undergo oxidation or reduction. Spectroscopically calibrated quantum-chemical computations show that the complex has high-spin iron(II) in a pseudotetrahedral coordination environment, which features interactions with side chains of two histidines and a cysteine as well as the C=O of Gly45. In the (5)A(1) ground state, the d(z2) orbital is doubly occupied. Mutation of Met121 to Ala leaves the metal site in a similar environment but creates a pocket for reversible binding of small anions to the iron(II) center. Specifically, azide forms a high-spin iron(II) complex and cyanide forms a low-spin iron(II) complex.

• 出版日期2012-12-5