6O10

E. coli cysteine desulfurase SufS

6O10 の概要

• エントリーDOI: 10.2210/pdb6o10/pdb
• 分子名称: Cysteine desulfurase, PYRIDOXAL-5'-PHOSPHATE, CHLORIDE ION, ... (4 entities in total)
• 機能のキーワード: cysteine desulfurase, sufs, persulfide, plp, transferase, lyase
• 由来する生物種: Escherichia coli (strain K12)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 44763.19

構造登録者

Dunkle, J.A.,Frantom, P.A. (登録日: 2019-02-17, 公開日: 2019-06-26, 最終更新日: 2023-10-11)

主引用文献

Blahut, M.,Wise, C.E.,Bruno, M.R.,Dong, G.,Makris, T.M.,Frantom, P.A.,Dunkle, J.A.,Outten, F.W.
Direct observation of intermediates in the SufS cysteine desulfurase reaction reveals functional roles of conserved active-site residues.
J.Biol.Chem., 294:12444-12458, 2019

PubMed Abstract: Iron-sulfur (Fe-S) clusters are necessary for the proper functioning of numerous metalloproteins. Fe-S cluster (Isc) and sulfur utilization factor (Suf) pathways are the key biosynthetic routes responsible for generating these Fe-S cluster prosthetic groups in Although Isc dominates under normal conditions, Suf takes over during periods of iron depletion and oxidative stress. Sulfur acquisition via these systems relies on the ability to remove sulfur from free cysteine using a cysteine desulfurase mechanism. In the Suf pathway, the dimeric SufS protein uses the cofactor pyridoxal 5'-phosphate (PLP) to abstract sulfur from free cysteine, resulting in the production of alanine and persulfide. Despite much progress, the stepwise mechanism by which this PLP-dependent enzyme operates remains unclear. Here, using rapid-mixing kinetics in conjunction with X-ray crystallography, we analyzed the pre-steady-state kinetics of this process while assigning early intermediates of the mechanism. We employed H123A and C364A SufS variants to trap Cys-aldimine and Cys-ketimine intermediates of the cysteine desulfurase reaction, enabling direct observations of these intermediates and associated conformational changes of the SufS active site. Of note, we propose that Cys-364 is essential for positioning the Cys-aldimine for Cα deprotonation, His-123 acts to protonate the Ala-enamine intermediate, and Arg-56 facilitates catalysis by hydrogen bonding with the sulfhydryl of Cys-aldimine. Our results, along with previous SufS structural findings, suggest a detailed model of the SufS-catalyzed reaction from Cys binding to C-S bond cleavage and indicate that Arg-56, His-123, and Cys-364 are critical SufS residues in this C-S bond cleavage pathway.

PubMed: 31248989
DOI: 10.1074/jbc.RA119.009471

実験手法

X-RAY DIFFRACTION (2 Å)