3VCP: The 2.2 Angstrom Structure Of Stc2 With Proline Bound In The Active Site

Citation:
Abstract
Herein, the structure resulting from in situ turnover in a chemically challenging quaternary ammonium oxidative demethylation reaction was captured via crystallographic analysis and analyzed via single-crystal spectroscopy. Crystal structures were determined for the Rieske-type monooxygenase, stachydrine demethylase, in the unliganded state (at 1.6 A resolution) and in the product complex (at 2.2 A resolution). The ligand complex was obtained from enzyme aerobically cocrystallized with the substrate stachydrine (N,N-dimethylproline). The ligand electron density in the complex was interpreted as proline, generated within the active site at 100 K by the absorption of X-ray photon energy and two consecutive demethylation cycles. The oxidation state of the Rieske iron-sulfur cluster was characterized by UV-visible spectroscopy throughout X-ray data collection in conjunction with resonance Raman spectra collected before and after diffraction data. Shifts in the absorption band wavelength and intensity as a function of absorbed X-ray dose demonstrated that the Rieske center was reduced by solvated electrons generated by X-ray photons; the kinetics of the reduction process differed dramatically for the liganded complex compared to unliganded demethylase, which may correspond to the observed turnover in the crystal.
PDB ID: 3VCPDownload
MMDB ID: 97126
PDB Deposition Date: 2012/1/4
Updated in MMDB: 2012/02
Experimental Method:
x-ray diffraction
Resolution: 2.2  Å
Source Organism:
Similar Structures:
Biological Unit for 3VCP: trimeric; determined by author and by software (PISA)
Molecular Components in 3VCP
Label Count Molecule
Proteins (3 molecules)
3
Ring-hydroxylating Dioxygenase
Molecule annotation
Chemicals (9 molecules)
1
3
2
3
3
3
* Click molecule labels to explore molecular sequence information.

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