Peptides from purified soybean β-conglycinin inhibit fatty acid synthase by interaction with the thioesterase catalytic domain

Cristina Martinez-Villaluenga, Sanjeewa G. Rupasinghe, Mary A. Schuler, Elvira Gonzalez De Mejia

Research output: Contribution to journalArticle

Abstract

Fatty acid synthase (FAS) is uniquely expressed at high levels in cancer cells and adipose tissue. The objectives of this study were to identify, purify and validate soy FAS inhibitory peptides and to predict their binding modes. Soy peptides were isolated from hydrolysates of purified β-conglycinin by co-immunoprecipitation and identified using LC-MS/MS. Three peptides, KNPQLR, EITPEKNPQLR and RKQEEDEDEEQQRE, inhibited FAS. The biological activity of these peptides was confirmed by their inhibitory activity against purified chicken FAS (IC50 = 79, 27 and 16 μm, respectively) and a high correlation (r = -0.7) with lipid accumulation in 3T3-L1 adipocytes. The FAS inhibitory potency of soy peptides also correlated with their molecular mass, pI value and the number of negatively charged and hydrophilic residues. Molecular modeling predicted that the large FAS inhibitory peptides (EITPEKNPQLR and RKQEEDEDEEQQRE) bond to the thioesterase domain of human FAS with lower interaction energies (-442 and -353 kcal·mol-1, respectively) than classical thioesterase inhibitors (Orlistat, -91 kcal·mol -1 and C75, -51 kcal·mol-1). Docking studies suggested that soy peptides blocked the active site through interactions within the catalytic triad, the interface cavity and the hydrophobic groove in the human FAS thioesterase domain. FAS thioesterase inhibitory activities displayed by the synthetic soy peptides EITPEKNPQLR and RKQEEDEDEEQQRE (IC50 = 10.1 ± 1.6 and 10.7 ± 4.4 μm, respectively) were higher than C75 (58.7 μm) but lower than Orlistat (0.9 μm). This is the first study to identify FAS inhibitory peptides from purified β-conglycinin hydrolysates and predict their binding modes at the molecular level, leading to their possible use as nutraceuticals.

Original languageEnglish (US)
Pages (from-to)1481-1493
Number of pages13
JournalFEBS Journal
Volume277
Issue number6
DOIs
StatePublished - Mar 1 2010

Fingerprint

Fatty Acid Synthases
Soybeans
Catalytic Domain
Peptides
Inhibitory Concentration 50
Molecular modeling
Molecular mass
Dietary Supplements
Bioactivity
Immunoprecipitation
Adipocytes
Adipose Tissue
Chickens
Cells
Tissue
Lipids

Keywords

  • Fatty acid synthase
  • Inhibitors
  • Soybean
  • Thioesterase
  • β-conglycinin-derived peptides

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Peptides from purified soybean β-conglycinin inhibit fatty acid synthase by interaction with the thioesterase catalytic domain. / Martinez-Villaluenga, Cristina; Rupasinghe, Sanjeewa G.; Schuler, Mary A.; Gonzalez De Mejia, Elvira.

In: FEBS Journal, Vol. 277, No. 6, 01.03.2010, p. 1481-1493.

Research output: Contribution to journalArticle

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AU - Rupasinghe, Sanjeewa G.

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AU - Gonzalez De Mejia, Elvira

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AB - Fatty acid synthase (FAS) is uniquely expressed at high levels in cancer cells and adipose tissue. The objectives of this study were to identify, purify and validate soy FAS inhibitory peptides and to predict their binding modes. Soy peptides were isolated from hydrolysates of purified β-conglycinin by co-immunoprecipitation and identified using LC-MS/MS. Three peptides, KNPQLR, EITPEKNPQLR and RKQEEDEDEEQQRE, inhibited FAS. The biological activity of these peptides was confirmed by their inhibitory activity against purified chicken FAS (IC50 = 79, 27 and 16 μm, respectively) and a high correlation (r = -0.7) with lipid accumulation in 3T3-L1 adipocytes. The FAS inhibitory potency of soy peptides also correlated with their molecular mass, pI value and the number of negatively charged and hydrophilic residues. Molecular modeling predicted that the large FAS inhibitory peptides (EITPEKNPQLR and RKQEEDEDEEQQRE) bond to the thioesterase domain of human FAS with lower interaction energies (-442 and -353 kcal·mol-1, respectively) than classical thioesterase inhibitors (Orlistat, -91 kcal·mol -1 and C75, -51 kcal·mol-1). Docking studies suggested that soy peptides blocked the active site through interactions within the catalytic triad, the interface cavity and the hydrophobic groove in the human FAS thioesterase domain. FAS thioesterase inhibitory activities displayed by the synthetic soy peptides EITPEKNPQLR and RKQEEDEDEEQQRE (IC50 = 10.1 ± 1.6 and 10.7 ± 4.4 μm, respectively) were higher than C75 (58.7 μm) but lower than Orlistat (0.9 μm). This is the first study to identify FAS inhibitory peptides from purified β-conglycinin hydrolysates and predict their binding modes at the molecular level, leading to their possible use as nutraceuticals.

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