Isopentenyl pyrophosphate c5 pp

Isopentenyl pyrophosphate (C₅-PP) and farnesyl pyrophosphate (C₁₅-PP) were purchased from Sigma. [¹⁴C]C₅-PP (50.6 mCi/mmol) was purchased from Perkin Elmer Life Sciences. The [¹⁴C]C₅₅-PP substrate was prepared as described below by successive condensations of [¹⁴C]C₅-PP onto C₁₅-PP catalysed by the purified UppS enzyme. The n-dodecyl-β-D-maltopyranoside (DDM) detergent was purchased from Anatrace and nickel-nitrilotriacetate-agarose (Ni²⁺-NTA-agarose) was from Qiagen. Compounds were purchased from Enamine and ChemBridge, and their identity and purity were confirmed (see Supplementary Information). All other materials were reagent grade and obtained from commercial sources.

Undecaprenyl-pyrophosphate (C₅₅-PP) and undecaprenyl-phosphate (C₅₅-P) were provided by the Institute of Biochemistry and Biophysics of the Polish Academy of Sciences. Farnesyl-pyrophosphate (C₁₅-PP) and isopentenyl-pyrophosphate (C₅-PP) were from Sigma and [¹⁴C]C₅-PP was purchased from Perkin Elmer Life Sciences. Diacylglycerol-pyrophosphate was from Avanti Polar Lipids and phosphatidylglycerol-phosphate was synthesized by using purified E. coli PgsA enzyme, essentially as described by Lu et al. [19 (link)]. n-dodecyl-β-D-maltoside (DDM) was purchased from Anatrace and isopropyl-β-D-thiogalactopyranoside (IPTG) from Eurogentec. C₅₅-PP synthase UppS was purified as described before [8 (link)]. DNA ligase and restriction enzymes were obtained from New England Biolabs, and DNA purification kits were from Promega and Macherey-Nagel. Synthesis of oligonucleotides and DNA sequencing were performed by Eurofins-MWG. All other materials were reagent grade and obtained from commercial sources.