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| Content Provider | PubMed Central |
|---|---|
| Author | Felix, Rohdich Ferdinand, Zepeck Adam, Petra Hecht, Stefan Kaiser, Johannes Ralf, Laupitz Tobias, Gräwert Sabine, Amslinger Eisenreich, Wolfgang Bacher, Adelbert Arigoni, Duilio |
| Copyright Year | 2003 |
| Abstract | Earlier in vivo studies have shown that the sequentialaction of the IspG and IspH proteins is essential for the reductivetransformation of 2C-methyl-d-erythritol2,4-cyclodiphosphate into dimethylallyl diphosphate andisopentenyl diphosphate via1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate. Arecombinant fusion protein comprising maltose binding protein and IspGprotein domains was purified from a recombinant Escherichiacoli strain. The purified protein failed to transform2C-methyl-d-erythritol 2,4-cyclodiphosphateinto 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate, butcatalytic activity could be restored by the addition of crude cellextract from an ispG-deficient E. coli mutant.This indicates that auxiliary proteins are required, probably asshuttles for redox equivalents. On activation by photoreduced10-methyl-5-deaza-isoalloxazine, the recombinant protein catalyzed theformation of 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphatefrom 2C-methyl-d-erythritol2,4-cyclodiphosphate at a rate of 1nmol⋅min−1⋅mg−1. Similarly,activation by photoreduced 10-methyl-5-deaza-isoalloxazine enabledpurified IspH protein to catalyze the conversion of1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate into a 6:1mixture of isopentenyl diphosphate and dimethylallyl diphosphate at arate of 0.4 μmol⋅min−1⋅mg−1. IspHprotein could also be activated by a mixture of flavodoxin, flavodoxinreductase, and NADPH at a rate of 3nmol⋅min−1⋅mg−1. The strikingsimilarities of IspG and IspH protein are discussed, and plausiblemechanistic schemes are proposed for the two reactions. |
| Related Links | http://dx.doi.org/10.1073/pnas.0337742100 |
| Ending Page | 1591 |
| Page Count | 6 |
| Starting Page | 1586 |
| File Format | |
| ISSN | 00278424 |
| e-ISSN | 10916490 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Issue Number | 4 |
| Volume Number | 100 |
| Language | English |
| Publisher | National Academy of Sciences |
| Publisher Date | 2003-02-18 |
| Access Restriction | Open |
| Rights Holder | National Academy of Sciences |
| Subject Keyword | General Research in Higher Education |
| Content Type | Text |
| Resource Type | Article |
| Subject | Multidisciplinary |
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