| Autor: |
Mason J, Appel, Katlyn K, Meier, Julien, Lafrance-Vanasse, Hyeongtaek, Lim, Chi-Lin, Tsai, Britt, Hedman, Keith O, Hodgson, John A, Tainer, Edward I, Solomon, Carolyn R, Bertozzi |
| Rok vydání: |
2019 |
| Předmět: |
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| Zdroj: |
Proceedings of the National Academy of Sciences of the United States of America. 116(12) |
| ISSN: |
1091-6490 |
| Popis: |
The formylglycine-generating enzyme (FGE) is required for the posttranslational activation of type I sulfatases by oxidation of an active-site cysteine to C(α)-formylglycine. FGE has emerged as an enabling biotechnology tool due to the robust utility of the aldehyde product as a bioconjugation handle in recombinant proteins. Here, we show that Cu(I)–FGE is functional in O(2) activation and reveal a high-resolution X-ray crystal structure of FGE in complex with its catalytic copper cofactor. We establish that the copper atom is coordinated by two active-site cysteine residues in a nearly linear geometry, supporting and extending prior biochemical and structural data. The active cuprous FGE complex was interrogated directly by X-ray absorption spectroscopy. These data unambiguously establish the configuration of the resting enzyme metal center and, importantly, reveal the formation of a three-coordinate tris(thiolate) trigonal planar complex upon substrate binding as furthermore supported by density functional theory (DFT) calculations. Critically, inner-sphere substrate coordination turns on O(2) activation at the copper center. These collective results provide a detailed mechanistic framework for understanding why nature chose this structurally unique monocopper active site to catalyze oxidase chemistry for sulfatase activation. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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