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Methylerythritol Phosphate Pathway: Enzymatic Evidence for a Rotation in the LytB/IspH‐Catalyzed Reaction
Author(s) -
Chaig Philippe,
Petit Benoît Eric,
Vincent Bruno,
Allouche Lionel,
Seemann Myriam
Publication year - 2020
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.201904676
Subject(s) - chemistry , enzyme , substrate (aquarium) , stereochemistry , catalysis , phosphate , biosynthesis , biochemistry , oceanography , geology
IspH/LytB, an oxygen‐sensitive [4Fe‐4S] enzyme, catalyzes the last step of the methylerythritol phosphate (MEP) pathway, a target for the development of new antimicrobial agents. This metalloenzyme converts ( E )‐4‐hydroxy‐3‐methylbut‐2‐en‐1‐yl diphosphate (HMBPP) into the two isoprenoid precursors: isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP). Here, the synthesis of ( S )‐[4‐ 2 H 1 ]HMBPP and ( R )‐[4‐ 2 H 1 ]HMBPP is reported together with a detailed NMR analysis of the products formed after their respective incubation with E. coli IspH/LytB in the presence of the biological reduction system used by E. coli to reduce the [4Fe‐4S] center. ( S )‐[4‐ 2 H 1 ]HMBPP was converted into [4‐ 2 H 1 ]DMAPP and ( E )‐[4‐ 2 H 1 ]IPP, whereas ( R )‐[4‐ 2 H 1 ]HMBPP yielded [4‐ 2 H 1 ]DMAPP and ( Z )‐[4‐ 2 H 1 ]IPP, hence providing the direct enzymatic evidence that the mechanism catalyzed by IspH/LytB involves a rotation of the CH 2 OH group of the substrate to display it away from the [4Fe‐4S].