Acquisition and biosynthesis of saturated and unsaturated fatty acids by trypanosomatids

Uttaro, Antonio D.

doi:10.1016/j.molbiopara.2014.04.001

Cited by 29 publications

(38 citation statements)

References 80 publications

(126 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…It has been previously proposed that these differences could be due to culture conditions, the age of the culture and the origin of the Leishmania strains [25]. Previous studies reported that both ∆6 and ∆5 elongases and ∆6, ∆5 and ∆4 desaturases were expressed in Leishmania major for the complete pathway of polyunsaturated fatty acid biosynthesis [1,31]. Accordingly, both n-3 and n-6 elongation/desaturation products were detected in both L. donovani and L. infantum.…”

Section: Discussionmentioning

confidence: 99%

“…Phospholipids are believed to be critical to virulence and viability in Leishmania [8]. As components of phospholipids and glycosylphosphatidylinositol anchors, fatty acids are responsible for forming the core of biological membranes and the correct localization of proteins within membranes [1]. The high phosphorylation dynamics during promastigote to amastigote differentiation [33] also suggests that protein phosphorylation cascades are involved in the developmental regulation of Leishmania parasites.…”

Section: Discussionmentioning

confidence: 99%

“…Recent interest has grown for the role of lipids in Trypanosomatids and especially in Trypanosoma cruzi, T. brucei and Leishmania spp. [1]. Lipid metabolism is of paramount importance for parasites, and especially for intracellular parasites, which rely on a complex system of uptake and synthesis mechanisms to satisfy their lipid needs.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Changes in Lipid and Fatty Acid Composition During Intramacrophagic Transformation of Leishmania donovani Complex Promastigotes into Amastigotes

Messaoud

Guichard

Lawton

et al. 2017

Lipids

View full text Add to dashboard Cite

Leishmania sp., are trypanosomatid parasites that are phagocytized by human and animal macrophages. Transformation from the vector promastigote stage to the intracellular amastigote host cell stage is mandatory, since development in the host depends on the internalization of the parasite. We identified and analyzed the lipids involved in the promastigote to amastigote transformation process in the Leishmania donovani complex. Four lipid classes, phospholipids, free fatty acids, triglycerides and sterols were studied. The derivatization method of Bligh and Dyer was used to establish the fatty acid composition in each stage of the parasite. To stay within the context of Leishmania infection, we used amastigotes extracted from macrophages after experimental in vitro infection. The purification process was checked by electronic microscopy, the absence of major contamination by host-cell debris and a correct purification yield validated our experimental model. Our results show that free fatty acids and cholesterol increased, whereas triglycerides and ergosterol decreased during the transition between promastigotes to amastigotes. With respect to phospholipid classes, we found increased proportion of sphingomyelin and phosphatidylserine and lowered proportion of phosphatidylinositol and lysophosphatidylethanolamine. Regarding fatty acid composition, a significant increase of n-7 fatty acids was observed in amastigotes. Overall, the total n-6 fatty acids were decreased in PL. Several of the changes were also observed in TG and free fatty acids. Particularly, n-7 fatty acids and 20:4n-6 were highly increased, whereas n-9 fatty acid and n-6 precursors decreased.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Changes in Lipid and Fatty Acid Composition During Intramacrophagic Transformation of Leishmania donovani Complex Promastigotes into Amastigotes

Messaoud

Guichard

Lawton

et al. 2017

Lipids

View full text Add to dashboard Cite

show abstract

“…Trypanosoma brucei is the causative agent of the sleeping sickness in Africa, whereas Trypanosoma cruzi causes Chagas' disease, the American trypanosomiasis. They belong to the class Kinetoplastea, which comprises organisms extensively studied due to their impact on human and animal health (Barrett et al 2003) and because of the remarkable biological characteristics they have (Ginger 2005), one of which being related to the fatty acid (FA) biosynthesis (Uttaro 2014). …”

Section: Introductionmentioning

confidence: 99%

Biosynthesis of very long chain fatty acids in Trypanosoma cruzi

Livore

Uttaro

2014

Parasitol Res

Self Cite

View full text Add to dashboard Cite

Trypanosoma brucei and Trypanosoma cruzi showed similar fatty acid (FA) compositions, having a high proportion of unsaturated FAs, mainly 18:2Δ9,12 (23-39%) and 18:1Δ9 (11-17%). C22 polyunsaturated FAs are in significant amounts only in T. brucei (12-20%) but represent a mere 2% of total FAs in T. cruzi. Both species have also similar profiles of medium- and long-chain saturated FAs, from 14:0 to 20:0. Interestingly, procyclic and bloodstream forms of T. brucei lack very long chain FAs (VLCFAs), whereas epimastigotes and trypomastigotes of T. cruzi contain 22:0 (0.1-0.2%), 24:0 (1.5-2%), and 26:0 (0.1-0.2%). This is in agreement with the presence of an additional FA elongase gene (TcELO4) in T. cruzi. TcELO4 was expressed in a Saccharomyces cerevisiae mutant lacking the endogenous ScELO3, rescuing the synthesis of saturated and hydroxylated C26 FAs in the yeast. Expression of TcELO4 also rescued the synthetic lethality of a ScELO2, ScELO3 double mutation, and the VLCFA profile of the transformed yeast was similar to that found in T. cruzi. By identifying TcELO4 as the enzyme responsible for the elongation of FA from 16:0 and 18:0 up to 26:0, with 24:0 being the preferred product, this work completed the characterization of FA elongases in Trypanosoma spp.

show abstract

“…This has led us to deal with the expression in yeast of trypanosomatid elongases and desaturases which required the use of PUFA like linolenic acids (18:3), AA, docosapentaenoic acid (DPA), or docosatetraenoic acid (DTA) as substrates. [6][7][8] The low incorporation of several of these PUFA have hampered the complete characterization of some enzymes, as the corresponding products usually appeared at the limit of detection, even using sensitive technologies like gas chromatography coupled to mass spectrometry (GCMS). Trypanosomatids also contain multiple ACS with different substrate specificities.…”

Section: Introductionmentioning

confidence: 99%

Improved Characterization of Polyunsaturated Fatty Acids Desaturases and Elongases by Co‐Expression in Saccharomyces cerevisiae with a Protozoan Acyl‐CoA Synthetase

Tripodi

Berardi

Uttaro

2018

Euro J Lipid Sci & Tech

Self Cite

View full text Add to dashboard Cite

Saccharomyces cerevisiae is a valuable host for the expression and characterization of eukaryotic enzymes involved in polyunsaturated fatty acid (PUFA) biosynthesis, such as elongases and desaturases. The yeast allows a correct subcellular localization of these proteins, provides electron donors required by desaturases and is unable to synthesize PUFA that could interfere in the enzymes characterization. Unfortunately, S. cerevisiae incorporates very long chain PUFAs inefficiently, which could interfere in the characterization of enzymes using these substrates. Acyl‐CoA synthetases (ACS) are involved in fatty acids uptake, and catalyze the synthesis of the corresponding CoA thioesters. ACS provides the substrates for elongases, acyl‐CoA desaturases and acyl transferases. Transferases are required to synthesize phospholipids which in turn, are substrates for acyl‐lipid desaturases. Expression in yeast of Trypanosoma brucei ACS1 notably improves the uptake of a wide variety of PUFA. Co‐expression of ACS1 with Elo5 elongase from Leishmania major or Des4 desaturase from T. brucei showes, respectively, 2‐ and 5.6‐fold increases in the uptake of the PUFA substrates and 2.4‐ and 3.5‐fold increases in substrate conversion. It also allows to produce significant amount of Des4 desaturase product for further analysis, whereas it is obtained in trace amounts when the enzyme is expressed alone. Practical applications: In this report, the use of yeast strains expressing ACS1 is proposed as a useful tool in the characterization of polyunsaturated fatty acids desaturases and elongases. Furthermore, this model could be used for the production of nutraceutical PUFA. S. cerevisiae (Sc) expressing an Acyl‐CoA synthetase from T. brucei (ACS1) significantly increases the intake of polyunsaturated fatty acids (PUFA), which are unusual for the yeast. It improves the possibility to study enzymes from the PUFA synthetic pathway, such as desaturases and elongases, by co‐expression in a Sc‐ACS1 background.

show abstract

Acquisition and biosynthesis of saturated and unsaturated fatty acids by trypanosomatids

Cited by 29 publications

References 80 publications

Changes in Lipid and Fatty Acid Composition During Intramacrophagic Transformation of Leishmania donovani Complex Promastigotes into Amastigotes

Changes in Lipid and Fatty Acid Composition During Intramacrophagic Transformation of Leishmania donovani Complex Promastigotes into Amastigotes

Biosynthesis of very long chain fatty acids in Trypanosoma cruzi

Improved Characterization of Polyunsaturated Fatty Acids Desaturases and Elongases by Co‐Expression in Saccharomyces cerevisiae with a Protozoan Acyl‐CoA Synthetase

Contact Info

Product

Resources

About