Short-chain polyisoprenoids in the yeast Saccharomyces cerevisiae — New companions of the old guys

Abstract

Dolichols are, among others, obligatory cofactors of protein glycosylation in eukaryotic cells. It is well known that yeast cells accumulate a family of dolichols with Dol-15/16 dominating while upon certain physiological conditions a second family with Dol-21 dominating is noted. In this report we identified the presence of additional short-chain length polyprenols - all-trans Pren-7 in three yeast strains (SS328, BY4741 and L5366), Pren-7 was accompanied by traces of putative Pren-6 and -8. Moreover, in two of these strains a single polyprenol mainly-cis-Pren-11 was synthesized at the stationary phase of growth. Identity of polyprenols was confirmed by HR-HPLC/MS, NMR and metabolic labeling. Additionally, simvastatin inhibited their biosynthesis.

Poly-Saturated Dolichols from Filamentous Fungi Modulate Activity of Dolichol-Dependent Glycosyltransferase and Physical Properties of Membranes

“…An initial HPLC/UV analysis of polyprenols isolated from T. reesei revealed a complex mixture containing numerous additional compounds besides those found in mammalian or S. cerevisiae cells and eluting with retention times corresponding to Dol-18–Dol-20. This was confirmed by the finer liquid chromatography electrospray ionization-quadrupole-time of flight-mass spectrometry (LC-ESI-QTOF-MS, liquide chromatography electrospray ionization-quadrupole-time of flight-mass spectrometry) analysis again showing a much more complex mixture of compounds (Table 1, Figure S1) than that reported for S. cerevisiae (Dol-14 to Dol-18 with Dol-16 dominating) [5,6,7]. An LC-ESI-QTOF-MS analysis showed that the polyprenols from T. reesei comprise a complex mixture of compounds with 18, 19 or 20 isoprene units and an apparently different degree of saturation (Table 1, a full list is presented in Table S1).…”

“…The only dolichols from filamentous fungi described so far were those isolated from A. niger [11] and A. fumigatus [10], and rather unexpectedly they were found to differ structurally from typical dolichols, as exemplified by those from animals or S. cerevisiae . In addition to these structural differences, they were also longer than the yeast ones, which are up to 22 isoprene units long [5,6,7]. A. niger contained a family of exo-methylenehexahydroprenols of between 18 and 24 isoprene residues, and A. fumigatus had dolichols with two or three hydrogenated isoprenoid residues located at the ω-end.…”

“…The pattern of the dolichols varies between kingdoms. Thus, S. cerevisiae accumulates a two-family mixture of dolichols—from Dol-14 to Dol-18, with Dol-15 or Dol-16 dominating, and longer ones from Dol-19 to Dol-22, with Dol-21 dominating [5,6,7].…”

The history and recent advances in research of polyprenol and its derivatives

“…Since Tanaka presumed that the biosynthesis of natural rubber starts with an unidentified initiating species containing two trans-isoprene units and a peptide group [52], it is possible to assume that one intermediate in the biosynthetic pathway of rubber might be an epoxy compound of a rubber-like polyprenol. characterization of a C 55 polyprenol detected at the stationary phase of growth of some yeast strains indicated the presence of internal two trans-isoprene residues [41]. Pig liver also contains a family of shorter-chain polyprenols dominated by a C 55 -dolichol and a long chain (C 95-100 ) dolichol [42].…”

Long-Chain Polyisoprenoids Are Synthesized by AtCPT1 in Arabidopsis thaliana

“…Yeast polyisoprenoid: Yeast cells were harvested by centrifugation of 100 mL of yeast culture at 3300× g (Allegra; Beckman, Indianapolis, IN, USA) for 10 min at room temperature, washed with water and hydrolyzed for 1 h at 95 °C in 10 mL of solution containing 25% KOH and 65% ethanol supplemented with internal standard Pren-28 (10 μg). Subsequently, lipids were extracted with hexane, purified on silica gel columns, dissolved in propan-2-ol and analyzed as described earlier [36].…”