Abstract
Polar lipid biosynthesis in the thermoacidophilic archaeon Thermoplasma acidophilum was analyzed using terbinafine, an inhibitor of tetraether lipid biosynthesis. Cells of T. acidophilum were labeled with [14C]mevalonic acid, and their lipids were extracted and analyzed by two-dimensional thin-layer chromatography. Lipids labeled with [14C]mevalonic acid, [14C]glycerol, and [32P]orthophosphoric acid were extracted and hydrolyzed under different conditions to determine the structure of polar lipids. The polar lipids were estimated to be archaetidylglycerol, glycerophosphatidylcaldarchaetidylglycerol, caldarchaetidylglycerol, and β-l-gulopyranosylcaldarchaetidylglycerol, the main polar lipid of T. acidophilum. Pulse and chase experiments with terbinafine revealed that one tetraether lipid molecule is synthesized by head-to-head condensation of two molecules of archaetidylglycerol and that a sugar group of tetraether phosphoglycolipid is expected to attach to the tetraether lipid core after head-to-head condensation in T. acidophilum. A precursor accumulated in the presence of terbinafine with a fast-atom-bombardment mass spectrometry peak m/z 806 was compatible with archaetidylglycerol. The relative height of the peak m/z 806 decreased after removal of the inhibitor. The results suggest that most of the precursor, archaetidylglycerol, is in fully saturated form.
Similar content being viewed by others
References
Bligh EG, Dyer WJ (1959) A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37:911–917
Chen A, Poulter CD (1993) Purification and characterization of farnesyl diphosphate/geranylgeranyl diphosphate synthase. A thermostable bifunctional enzyme from Methanobacterium thermoautotrophicum. J Biol Chem 268:11002–11007
Chen A, Zhang D, Poulter CD (1993) (S)-geranylgeranylglyceryl phosphate synthase. Purification and characterization of the first pathway-specific enzyme in archaebacterial membrane lipid biosynthesis. J Biol Chem 268:21701–21705
De Rosa M, Gambacorta A, Nicolaus B, Chappe B, Albrecht P (1983) Isoprenoid ethers: backbone of complex lipids of the archaebacterium Sulfolobus solfataricus. Biochim Biophys Acta 753:249–256
Eguchi T, Takyo H, Morita M, Kakinuma K, Koga Y (2000) Unusual double-bond migration as a plausible key reaction in the biosynthesis of the isoprenoidal membrane lipids of methanogenic archaea. Chem Commun 1545–1546
Ekiel I, Sprott GD (1992) Identification of degradation artifacts formed upon treatment of hydroxydiether lipids from methanogens with methanolic HCl. Can J Microbiol 38:764–768
Koga Y, Nishihara M, Morii H, Akagawa-Matsushita M (1993) Ether polar lipids of methanogenic bacteria: structures, comparative aspects, and biosyntheses. Microbiol Rev 57:164–182
Kon T, Nemoto N, Oshima T, Yamagishi A (2002) Effects of a squalene epoxidase inhibitor, terbinafine, on ether lipid biosyntheses in a thermoacidophilic archaeon, Thermoplasma acidophilum. J Bacteriol 184:1395–1401
Kushwaha SC, Kates M, Sprott GD, Smith IC (1981) Novel polar lipids from the methanogen Methanospirillum hungatei GP1. Biochim Biophys Acta 664:156–173
Langworthy TA (1982) Lipids of Thermoplasma. Methods Enzymol 88:396–406
Langworthy TA, Smith, PF, Mayberry WR (1972) Lipids of Thermoplasma acidophilum. J Bacteriol 112:1193–1200
Moldoveanu N, Kates M (1988) Biosynthetic studies of the polar lipids of Halobacterium cutirubrum. Formation of isoprenyl ether intermediates. Biochim Biophys Acta 960:164–182
Morii H, Nishihara M, Koga Y (2000) CTP:2,3-di-O-geranylgeranyl-sn-glycero-1-phosphate cytidyltransferase in the methanogenic archaeon Methanothermobacter thermoautotrophicus. J Biol Chem 275:36568–36574
Nishihara M, Morii H, Koga Y (1989) Heptads of polar ether lipids of an archaebacterium, Methanobacterium thermoautotrophicum: Structure and biosynthetic relationship. Biochemistry 28:95–102
Ohnuma S, Suzuki M, Nishino T (1994) Archaebacterial ether-linked lipid biosynthetic gene. Expression cloning, sequencing, and characterization of geranylgeranyl-diphosphate synthase. J Biol Chem 269:14792–14797
Ross HMN, Grant WD (1985) Chemical methods in bacterial systematics. Wiley, New York
Ryder NS (1984) Microbial cell wall synthesis and autolysis. Elsevier, Amsterdam
Shimada H, Nemoto N, Shida Y, Oshima T, Yamagishi A (2002) Complete polar lipid composition of Thermoplasma acidophilum HO-62 determined by high-performance liquid chromatography with evaporative light-scattering detection. J Bacteriol 184:556–563
Soderberg T, Chen A, Poulter CD (2001) Geranylgeranylglyceryl phosphate synthase. Characterization of the recombinant enzyme from Methanobacterium thermoautotrophicum. Biochemistry 40:14847–14854
Sugai A, Uda I, Kon K, Ando S, Itoh YH, Itoh T (1996) Structural identification of minor phosphoinositol lipids in Sulfolobus acidocaldarius N-8. J Jpn Oil Chem Soc 45:327–333
Swain M, Brisson JR, Sprott GD, Cooper FP, Patel GB (1997) Identification of β-l-gulose as the sugar moiety of the main polar lipid Thermoplasma acidophilum. Biochim Biophys Acta 1345:56–64
Tachibana A (1994) A novel prenyltransferase, farnesylgeranyl diphosphate synthase, from the haloalkaliphilic archaeon, Natronobacterium pharaonis. FEBS Lett 341:291–294
Trincone A, De Rosa M, Gambacorta A, Lanzotti V, Nicolaus B, Harris JE, Grant WD (1988) A simple chromatographic procedure for the detection of cyclized archaebacterial glycerol-bisdiphytanyl-glycerol tetraether core lipids. J Gen Microbiol 134:3159–3163
Uda I, Sugai A, Kon K, Ando S, Itoh YH, Itoh T (1999) Isolation and characterization of novel neutral glycolipids from Thermoplasma acidophilum. Biochim Biophys Acta 1439:363–370
Uda I, Sugai A, Itoh YH, Itoh T (2000a) Characterization of caldarchaetidylglycerol analogs, dialkyl-type and trialkyl-type, from Thermoplasma acidophilum. Lipids 35:1155–1157
Uda I, Sugai A, Shimizu A, Itoh YH, Itoh T (2000b) Glucosylcaldarchaetidylglycerol, a minor phosphoglycolipid from Thermoplasma acidophilum. Biochim Biophys Acta 1484:83–86
Yasuda M, Oyaizu H, Yamagishi A, Oshima T (1995) Morphological variation of new Thermoplasma acidophilum isolates from Japanese hot springs. Appl Environ Microbiol 61:3482–3485
Zhang D, Poulter CD (1993) Biosynthesis of archaebacterial lipids in Halobacterium halobium and Methanobacterium thermoautotrophicum. J Org Chem 58:3919–3922
Acknowledgements
This work was supported by a Grant-in-Aid from the Ministry of Education, Culture, Sports, Science and Technology of Japan (09839033).
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by G. Antranikian
Rights and permissions
About this article
Cite this article
Nemoto, N., Shida, Y., Shimada, H. et al. Characterization of the precursor of tetraether lipid biosynthesis in the thermoacidophilic archaeon Thermoplasma acidophilum . Extremophiles 7, 235–243 (2003). https://doi.org/10.1007/s00792-003-0315-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00792-003-0315-x