title: LytB protein catalyzes the terminal step of the 2-C-methyl-D-erythritol-4-phosphate pathway of isoprenoid biosynthesis.
abstract: Recombinant LytB protein from the thermophilic eubacterium Aquifex aeolicus produced in Escherichia coli was purified to apparent homogeneity. The purified LytB protein catalyzed the reduction of (E)-4-hydroxy-3-methyl-but-2-enyl diphosphate (HMBPP) in a defined in vitro system. The reaction products were identified as isopentenyl diphosphate and dimethylallyl diphosphate. A spectrophotometric assay was established to determine the steady-state kinetic parameters of LytB protein. The maximal specific activity of 6.6+/-0.3 micromol x min(-1) x mg(-1) protein was determined at pH 7.5 and 60 degrees C. The k(cat) value of the LytB protein was 3.7+/-0.2 s(-1) and the K(m) value for HMBPP was 590+/-60 microM.
authors.affiliation: Jomaa Pharmaka GmbH, Frankfurter Strasse 50, 35392, Giessen, Germany. boran.altincicek@jomaa.de
authors.forename: Boran
authors.initials: B
authors.lastname: Altincicek
authors.forename: Evert C
authors.initials: EC
authors.lastname: Duin
authors.forename: Armin
authors.initials: A
authors.lastname: Reichenberg
authors.forename: Reiner
authors.initials: R
authors.lastname: Hedderich
authors.forename: Ann-Kristin
authors.initials: AK
authors.lastname: Kollas
authors.forename: Martin
authors.initials: M
authors.lastname: Hintz
authors.forename: Stefanie
authors.initials: S
authors.lastname: Wagner
authors.forename: Jochen
authors.initials: J
authors.lastname: Wiesner
authors.forename: Ewald
authors.initials: E
authors.lastname: Beck
authors.forename: Hassan
authors.initials: H
authors.lastname: Jomaa
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D002384:Catalysis; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D004896:Erythritol / Q000031:analogs & derivatives* / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins; D005051:Eubacterium / Q000235:genetics / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008956:Models, Chemical; D008969:Molecular Sequence Data; D010755:Organophosphates / Q000494:pharmacology; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization; D013403:Sugar Phosphates / Q000378:metabolism*; D013696:Temperature; D013997:Time Factors
chemical_list: C114232:2-C-methylerythritol 4-phosphate; C467061:4-hydroxy-3-methyl-2-butenyl diphosphate; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D010755:Organophosphates; D011994:Recombinant Proteins; D013403:Sugar Phosphates; D010088:Oxidoreductases; C109866:ispH protein, E coli; D004896:Erythritol