Interaction of bovine -lactalbumin with fatty acids as determined by partition equilibrium and fluorescence spectroscopy

Barbana, Chockry; Pérez, María D.; Sánchez, Lourdes; Dalgalarrondo, Michèle; Chobert, Jean‐Marc; Haertlé, Tomasz; Calvo, Miguel

doi:10.1016/j.idairyj.2005.01.007

Cited by 71 publications

(54 citation statements)

References 29 publications

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“…2. The spectrum of the holo form has two minima at 270 and 298 nm arising from spectral contribution of aromatic chromophores (tryptophan, tyrosine and phenylalanine) as previously reported [8,15]. In the presence of 20% ethanol, the spectrum is similar to that of the holo protein in the absence of ethanol (Fig.…”

Section: Influence Of Ethanol On the Structural Changes Of A-lactalbuminsupporting

confidence: 74%

“…It has been shown that a-lactalbumin binds 5-doxylstearic, stearic, palmitic and oleic acids and the binding affinity depends upon the protein state [8 -10]. Studies performed using fluorescence spectroscopy and partition equilibrium techniques indicated that the apo form of the bovine a-lactalbumin displays one binding site for oleic acid with an association constant of 4.6610 6 M -1 [8]. However, holo a-lactalbumin is unable to bind fatty acids [8,10].…”

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confidence: 99%

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Study of ethanol‐induced conformational changes of holo and apo α‐lactalbumin by spectroscopy and limited proteolysis

Wehbi¹,

Pérez²,

Dalgalarrondo³

et al. 2006

Molecular Nutrition Food Res

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This study was performed to contribute to the analysis of alpha-lactalbumin "molten globule" state by using spectral and proteolysis techniques. Samples of holo and apo alpha-lactalbumin in the presence of different concentrations of ethanol were analyzed. Results of fluorescence spectroscopy of both forms showed that as ethanol concentration increased, the tryptophanyl residues became more accessible to the solvent. Near circular dichroism spectra of holo alpha-lactalbumin indicated that its tertiary structure was maintained in 20% ethanol whereas it was altered in 30 and 40% ethanol. For apo alpha-lactalbumin, spectra were similar in all samples studied. Holo alpha-lactalbumin was resistant to trypsinolysis in 0% ethanol, whereas it was easily hydrolyzed in 20 and 30% ethanol. In the case of the apo form and in the absence of ethanol, 70% of the protein was degraded after 1 h. However, in the presence of 20 and 30% ethanol, the overall reaction rate was lowered. Peptides obtained after tryptic hydrolysis were identified by reversed-phase high-performance liquid chromatography coupled to mass spectrometry. Differences in population of produced peptides indicate the changes of folding intermediates present in the studied alpha-lactalbumin solutions. This study demonstrated that proteolytic enzymes are suitable tools to determine protein structure complementing physico-chemical studies.

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Section: Influence Of Ethanol On the Structural Changes Of A-lactalbuminsupporting

confidence: 74%

mentioning

confidence: 99%

Study of ethanol‐induced conformational changes of holo and apo α‐lactalbumin by spectroscopy and limited proteolysis

Wehbi¹,

Pérez²,

Dalgalarrondo³

et al. 2006

Molecular Nutrition Food Res

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“…For HAMLET, the stoichiometries so far reported thus ranged from less than 1 to 9 (the number of the OA mol-ecules per an HLA molecule) (20,(43)(44)(45), and the stoichiometries in HAMLET and GAMLET prepared by the present method were 8 and 7, respectively.…”

Section: Discussionmentioning

confidence: 67%

Molecular Mechanisms of the Cytotoxicity of Human α-Lactalbumin Made Lethal to Tumor Cells (HAMLET) and Other Protein-Oleic Acid Complexes

Nakamura

Aizawa

Kariya

et al. 2013

Journal of Biological Chemistry

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Background:The ␣-lactalbumin-oleic acid complex has a unique apoptotic activity for selectively killing tumor cells. Results: We identified the oleic acid-binding site in the human-and goat-␣-lactalbumin complexes by two-dimensional NMR. Conclusion: Oleic acid is bound to a loosely organized hydrophobic core of the proteins in the molten globule state. Significance: The results are crucial for understanding the molecular mechanisms of the cytotoxicity of the protein-oleic acid complexes.

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“…The strength of any association of fatty acid with the protein has to be sufficiently weak to allow the fatty acid be released from the protein to have a cytotoxic effect. Previous values reported in the literature would indicate that the association constant of OA to α-LA is relatively weak [28].…”

Section: Discussionmentioning

confidence: 83%

“…The equilibrium between a-LA and fatty acid is further complicated by the equilibrium between the fatty acid and column matrix. As the binding to α-LA has been reported to be quite weak [28], the absorption onto the column matrix may in fact limit the amount of fatty acid taken up by the protein. The variations in the affinity of different fatty acids for the column matrix and/or α-LA could be significant in the formation of the complexes using the column method.…”

Section: Discussionmentioning

confidence: 99%

The cytotoxicity of fatty acid/α‐lactalbumin complexes depends on the amount and type of fatty acid

Brinkmann

Brodkorb

Thiel

et al. 2013

Euro J Lipid Sci & Tech

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Complexes of the milk protein, α‐lactalbumin, and the fatty acid, oleic acid, have previously been shown to be cytotoxic. Complexes of α‐lactalbumin and five different fatty acids (vaccenic, linoleic, palmitoleic, stearic, and elaidic acid) were prepared and compared to those formed with oleic acid. All complexes were cytotoxic to human promyelocytic leukemia‐derived (HL‐60) cells but to different degrees depending on the fatty acid. The amount of fatty acid per α‐lactalbumin molecule was found to correlate with the cytotoxicity; the higher the number of fatty acids per protein, the more cytotoxic the complex. Importantly, all the tested fatty acids were also found to be cytotoxic on their own in a concentration dependent manner. The cytotoxic effect of complexes between α‐lactalbumin and linoleic acid, vaccenic acid, or oleic acid was further investigated using flow cytometry and found to induce cell death resembling apoptosis on Jurkat cells.Practical applications: Cytotoxic complexes of α‐lactalbumin and several different fatty acids could be produced. The cytotoxicity of all the variants is similar to that previously determined for α‐lactalbumin/oleic acid complexes.

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Interaction of bovine -lactalbumin with fatty acids as determined by partition equilibrium and fluorescence spectroscopy

Cited by 71 publications

References 29 publications

Study of ethanol‐induced conformational changes of holo and apo α‐lactalbumin by spectroscopy and limited proteolysis

Study of ethanol‐induced conformational changes of holo and apo α‐lactalbumin by spectroscopy and limited proteolysis

Molecular Mechanisms of the Cytotoxicity of Human α-Lactalbumin Made Lethal to Tumor Cells (HAMLET) and Other Protein-Oleic Acid Complexes

The cytotoxicity of fatty acid/α‐lactalbumin complexes depends on the amount and type of fatty acid

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