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

Gryz Gryz,Długoński Długoński,Bernat Bernat,Steczkiewicz Steczkiewicz,Perlińska-Lenart Perlińska-Lenart,Ginalski Ginalski,Swiezewska Swiezewska,Muszewska Muszewska,Lipko Lipko,Kruszewska Kruszewska,Jemiola-Rzeminska Jemiola-Rzeminska,Jozwiak Jozwiak,Gawarecka Gawarecka,Strzalka Strzalka,Piłsyk Piłsyk

doi:10.3390/ijms20123043

Abstract

Mono-saturated polyprenols (dolichols) have been found in almost all Eukaryotic cells, however, dolichols containing additional saturated bonds at the ω-end, have been identified in A. fumigatus and A. niger. Here we confirm using an LC-ESI-QTOF-MS analysis, that poly-saturated dolichols are abundant in other filamentous fungi, Trichoderma reesei, A. nidulans and Neurospora crassa, while the yeast Saccharomyces cerevisiae only contains the typical mono-saturated dolichols. We also show, using differential scanning calorimetry (DSC) and fluorescence anisotropy of 1,6-diphenyl-l,3,5-hexatriene (DPH) that the structure of dolichols modulates the properties of membranes and affects the functioning of dolichyl diphosphate mannose synthase (DPMS). The activity of this enzyme from T. reesei and S. cerevisiae was strongly affected by the structure of dolichols. Additionally, the structure of phosphatidylcholine (PC) and phosphatidylethanolamine (PE) model membranes was more strongly disturbed by the poly-saturated dolichols from Trichoderma than by the mono-saturated dolichols from yeast. By comparing the lipidome of filamentous fungi with that from S. cerevisiae, we revealed significant differences in the PC/PE ratio and fatty acids composition. Filamentous fungi differ from S. cerevisiae in the lipid composition of their membranes and the structure of dolichols. The structure of dolichols profoundly affects the functioning of dolichol-dependent enzyme, DPMS.

Highlights

Polyisoprenoid alcohols are found in all living organisms from bacteria to mammals
CPT catalyzes the elongation of the polyprenol chain by sequential addition of isopentenyl diphosphate (IPP) to a precursor farnesyl diphosphate (FPP) [2,3,4]
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

Summary

Introduction

Polyisoprenoid alcohols are found in all living organisms from bacteria to mammals They consist of up to more than 100 isoprene residues linked head-to-tail, with a hydroxyl group at one end (α-residue) and a hydrogen atom at the other (ω-end) [1]. In animals and yeast the double bond in the α-residue is hydrogenated, giving dolichols. In these organisms, dolichols, like all the other isoprenoids, are synthesized via the mevalonate pathway and the first dedicated enzyme of the polyprenol branch of this pathway is cis-prenyltransferase (dehydrodolichyl diphosphate synthase) (CPT), a key enzyme in dolichol synthesis. Eukaryotic cells synthesize polyisoprenoids with a specific number of isoprene residues forming a mixture of usually six to eight dolichol species of different chain lengths, while bacteria possess a single polyprenol, undecaprenol (Pren-11) [1]. 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]

Methods

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Discussion

Conclusion