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• W2024694051 abstract "(E)-4-Hydroxy-3-methylbut-2-enyl diphosphate reductase (IspH or LytB) catalyzes the terminal step of the MEP/DOXP pathway where it converts (E)-4-hydroxy-3-methylbut-2-enyl diphosphate (HMBPP) into the two products, isopentenyl diphosphate and dimethylallyl diphosphate. The reaction involves the reductive elimination of the C4 hydroxyl group, using a total of two electrons. Here we show that the active form of IspH contains a [4Fe-4S] cluster and not the [3Fe-4S] form. Our studies show that the cluster is the direct electron source for the reaction and that a reaction intermediate is bound directly to the cluster. This active form has been trapped in a state, dubbed FeS(A), that was detected by electron paramagnetic resonance (EPR) spectroscopy when one-electron-reduced IspH was incubated with HMBPP. In addition, three mutants of IspH have been prepared and studied, His42, His124, and Glu126 (Aquifex aeolicus numbering), with particular attention paid to the effects on the cluster properties and possible reaction intermediates. None of the mutants significantly affected the properties of the [4Fe-4S](+) cluster, but different effects were observed when one-electron-reduced forms were incubated with HMBPP. Replacing His42 led to an increased K(M) value and a much lower catalytic efficiency, confirming the role of this residue in substrate binding. Replacing the His124 also resulted in a lower catalytic efficiency. In this case, however, the enzyme showed the loss of the [4Fe-4S](+) EPR signal upon addition of HMBPP without the subsequent formation of the FeS(A) signal. Instead, a radical-type signal was observed in some of the samples, indicating that this residue plays a role in the correct positioning of the substrate. The incorrect orientation in the mutant leads to the formation of substrate-based radicals instead of the cluster-bound intermediate complex FeS(A). Replacing the Glu126 also resulted in a lower catalytic efficiency, with yet a third type of EPR signal being detected upon incubation with HMBPP. (31)P and (2)H ENDOR measurements of the FeS(A) species incubated with regular and (2)H-C4-labeled HMBPP reveal that the substrate binds to the enzyme in the proximity of the active-site cluster with C4 adjacent to the site of linkage between the FeS cluster and HMBPP. Comparison of the spectroscopic properties of this intermediate to those of intermediates detected in (E)-4-hydroxy-3-methylbut-2-enyl diphosphate synthase and ferredoxin:thioredoxin reductase suggests that HMBPP binds to the FeS cluster via its hydroxyl group instead of a side-on binding as previously proposed for the species detected in the inactive Glu126 variant. Consequences for the IspH reaction mechanism are discussed." @default.
• W2024694051 created "2016-06-24" @default.
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• W2024694051 date "2012-06-07" @default.
• W2024694051 modified "2023-09-25" @default.
• W2024694051 title "A Closer Look at the Spectroscopic Properties of Possible Reaction Intermediates in Wild-Type and Mutant (<i>E</i>)-4-Hydroxy-3-methylbut-2-enyl Diphosphate Reductase" @default.
• W2024694051 cites W1525488930 @default.
• W2024694051 cites W1538196126 @default.
• W2024694051 cites W1622339598 @default.
• W2024694051 cites W1967650766 @default.
• W2024694051 cites W1969243089 @default.
• W2024694051 cites W1970794882 @default.
• W2024694051 cites W1970815975 @default.
• W2024694051 cites W1972401663 @default.
• W2024694051 cites W1972430667 @default.
• W2024694051 cites W1972606544 @default.
• W2024694051 cites W1975131095 @default.
• W2024694051 cites W1976725431 @default.
• W2024694051 cites W1991907415 @default.
• W2024694051 cites W1992425917 @default.
• W2024694051 cites W1993818604 @default.
• W2024694051 cites W1995664405 @default.
• W2024694051 cites W1997893127 @default.
• W2024694051 cites W1999458818 @default.
• W2024694051 cites W2001437131 @default.
• W2024694051 cites W2003219159 @default.
• W2024694051 cites W2003306724 @default.
• W2024694051 cites W2005817850 @default.
• W2024694051 cites W2008872195 @default.
• W2024694051 cites W2009288340 @default.
• W2024694051 cites W2012382149 @default.
• W2024694051 cites W2014847912 @default.
• W2024694051 cites W2015946472 @default.
• W2024694051 cites W2028371684 @default.
• W2024694051 cites W2028539126 @default.
• W2024694051 cites W2038205004 @default.
• W2024694051 cites W2038675817 @default.
• W2024694051 cites W2038897702 @default.
• W2024694051 cites W2040005327 @default.
• W2024694051 cites W2041322046 @default.
• W2024694051 cites W2041852369 @default.
• W2024694051 cites W2043636038 @default.
• W2024694051 cites W2047520039 @default.
• W2024694051 cites W2062737545 @default.
• W2024694051 cites W2063949270 @default.
• W2024694051 cites W2068066782 @default.
• W2024694051 cites W2075922180 @default.
• W2024694051 cites W2080172904 @default.
• W2024694051 cites W2082094739 @default.
• W2024694051 cites W2089119194 @default.
• W2024694051 cites W2095445547 @default.
• W2024694051 cites W2103243692 @default.
• W2024694051 cites W2108177339 @default.
• W2024694051 cites W2128197293 @default.
• W2024694051 cites W2128951621 @default.
• W2024694051 cites W2133350645 @default.
• W2024694051 cites W2134761247 @default.
• W2024694051 cites W2144876507 @default.
• W2024694051 cites W2145741655 @default.
• W2024694051 cites W2146684384 @default.
• W2024694051 cites W2151486064 @default.
• W2024694051 cites W2164267446 @default.
• W2024694051 cites W2168059482 @default.
• W2024694051 cites W2272525520 @default.
• W2024694051 cites W2288883598 @default.
• W2024694051 cites W2325002332 @default.
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• W2024694051 doi "https://doi.org/10.1021/bi3001215" @default.
• W2024694051 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/3426640" @default.
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