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Fetal Bovine Serum Concentration Affects Δ9 Desaturase Activity of Trypanosoma cruzi

  • Original Article
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Lipids

Abstract

Fetal bovine serum (FBS) is an important factor in the culture of Trypanosoma cruzi, since this parasite obtains and metabolizes fatty acids (FAs) from the culture medium, and changes in FBS concentration reduce the degree of unsaturation of FAs in phosphoinositides. When T. cruzi epimastigotes were cultured with 5% instead of 10% FBS, and stearic acid was used as the substrate, ∆9 desaturase activity decreased by 50%. Apparent K m and V m values for stearic acid, determined from Lineaweaver–Burk plots, were 2 μM and 219 pmol/min/mg of protein, respectively. In studies of the requirement for reduced pyridine nucleotide, ∆9 desaturase activity reached a maximum with 8 μM NADH and then remained constant; the apparent K m and V m were 4.3 μM and 46.8 pmol/min/mg of protein, respectively. The effect of FBS was observed only for ∆9 desaturase activity; ∆12 desaturase activity was not affected. The results suggest that decreased FBS in culture medium is a signal that modulates ∆9 desaturase activity in T. cruzi epimastigotes.

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Abbreviations

ATP:

Adenosine triphosphate

FBS:

Fetal bovine serum

EDTA:

Ethylenediaminetetraacetic acid

FAME:

Fatty acid methyl esters

FFA:

Free fatty acid

FAs:

Fatty acids

HPLC:

High performance liquid chromatography

HEPES:

(4-(2-Hydroxyethyl)-1-piperazineethanesulfonic acid

NADH:

Nicotinamide adenine dinucleotide

NADPH:

Nicotinamide adenine dinucleotide phosphate

NL:

Neutral lipid

PL:

Phospholipid

PMSF:

Phenylmethylsulphonyl fluoride

PUFAs:

Polyunsaturated fatty acids

PC:

Phosphatidylcholine

SEM:

Standard error of the mean

TLC:

Thin layer chromatography

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Acknowledgments

The authors thank Dr. Marta Aveldaño and Dr. Carlos Marra for determination of FBS FA composition, and FA analysis, respectively. This work was supported by FONCyT, Argentina and SECyT, UNRC, Río Cuarto, Córdoba, Argentina. A.L.V. is a research career scientist of CONICET, Argentina.

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Authors and Affiliations

  1. Departamento de Biología Molecular, Facultad de Ciencias Exactas, Físico-Químicas y Naturales, Universidad Nacional de Río Cuarto, CPX5804BYA, Río Cuarto, Córdoba, Argentina

    Ana L. Villasuso, Patricio Romero, Mariela Woelke, Patricia Moyano, Estela Machado & Mirta García de Lema

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  1. Ana L. Villasuso
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  2. Patricio Romero
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  6. Mirta García de Lema
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Correspondence to Mirta García de Lema.

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Villasuso, A.L., Romero, P., Woelke, M. et al. Fetal Bovine Serum Concentration Affects Δ9 Desaturase Activity of Trypanosoma cruzi . Lipids 45, 275–283 (2010). https://doi.org/10.1007/s11745-010-3387-2

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  • DOI: https://doi.org/10.1007/s11745-010-3387-2

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