6MUJ

Formylglycine generating enzyme bound to copper

Summary for 6MUJ

• Entry DOI: 10.2210/pdb6muj/pdb
• Descriptor: Formylglycine-generating enzyme, CALCIUM ION, COPPER (II) ION, ... (9 entities in total)
• Functional Keywords: formylglycine, copper oxidase, metalloenzyme, metal binding protein
• Biological source: Streptomyces coelicolor
• Total number of polymer chains: 5
• Total formula weight: 173784.36

Authors

Lafrance-Vanasse, J.,Appel, M.J.,Tsai, C.-L.,Bertozzi, C.,Tainer, J.A. (deposition date: 2018-10-23, release date: 2019-02-27, Last modification date: 2023-10-11)

Primary citation

Appel, M.J.,Meier, K.K.,Lafrance-Vanasse, J.,Lim, H.,Tsai, C.L.,Hedman, B.,Hodgson, K.O.,Tainer, J.A.,Solomon, E.I.,Bertozzi, C.R.
Formylglycine-generating enzyme binds substrate directly at a mononuclear Cu(I) center to initiate O2activation.
Proc. Natl. Acad. Sci. U.S.A., 116:5370-5375, 2019

PubMed Abstract: The formylglycine-generating enzyme (FGE) is required for the posttranslational activation of type I sulfatases by oxidation of an active-site cysteine to C-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 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 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.

PubMed: 30824597
DOI: 10.1073/pnas.1818274116

Experimental method

X-RAY DIFFRACTION (2.249 Å)