Effect of temperature/ethanol on the secondary structure of bovine Apo Alpha-Lactalbumin investigated by FTIR/2D IR correlation spectoscropy
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Notice bibliographique
Résumé
Whey proteins play a vital role in the manufacture of food products due to their nutritional value and versatile functional properties.-Lactalbumin (-LA) is the second most abundant protein in bovine whey and the most abundant protein of human whey.-LA is a low-molecular-weight (14.2 kDa) and acidic (pI 4-5) protein that is produced in the lactating mammary glands and has a role in lactose biosynthesis.BAMLET/HAMLET (bovine/human alpha-lactalbumin made lethal to tumor cells) are complexes of -LA and oleic acid that have been shown to have cytotoxic effects on tumor cells but not on healthy cells.In vitro, it has been reported that BAMLET-type complexes can be prepared by heating a solution of bovine apo (calciumdepleted) -LA in sodium phosphate buffer to which an ethanol solution of oleic acid has been added.However, the possibility that the presence of ethanol may facilitate the complexation of oleic acid with -LA by affecting the thermal denaturation of the protein has not been investigated In the present study, the combined effects of ethanol and temperature on the secondary structure of bovine apo -LA were examined by variable-temperature Fourier transform infrared (VT-FTIR) spectroscopy in conjunction with Fourier self-deconvolution (a resolution enhancement technique) and two-dimensional cross correlation spectroscopy (2D CCS).At room temperature, an increase in -helical and -structure content at the expense of 3 10 -helices and turns was observed as a function of increasing the concentration of ethanol (from ~2.5 to 33% w/v).These findings are consistent with the fluorescence and proteolysis studies of -LA reported in the literature, which showed a similar effect of ethanol on the secondary structure of -LA.Subjecting bovine apo -LA solutions to a heating-cooling cycle (heating from 25 to 95 0 C and cooling from 95 to 25 0 C) in the presence of varying concentrations of ethanol was found to alter the protein's secondary structure.At any concentration of ethanol the -helix and 3 10 -helices of the secondary structure of bovine apo -LA were lost upon heating of the protein.The sequences of the changes in secondary structure during the heating and cooling cycles were elucidated by 2D CCS.The results revealed that the protein refolded during the cooling cycle by reversal of the sequence of unfolding events during the heating cycle only in the presence of 20% or higher concentration of ethanol.Overall, the present study supported the ethanol-induced reversible thermal denaturation of bovine apo--LA.
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Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Apprise à partir de 10 348 étiquettes directes de Codex et de 10 348 étiquettes directes de Gemma. Le mode candidate est l'union des têtes enseignantes seuillées; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont ni des étiquettes humaines ni des étiquettes directes de modèles de pointe.
Scores Codex et Gemma par catégorie
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,002 | 0,002 |
| Méta-épidémiologie (sens strict) | 0,001 | 0,001 |
| Méta-épidémiologie (sens large) | 0,002 | 0,000 |
| Bibliométrie | 0,000 | 0,001 |
| Études des sciences et des technologies | 0,001 | 0,000 |
| Communication savante | 0,000 | 0,000 |
| Science ouverte | 0,001 | 0,000 |
| Intégrité de la recherche | 0,002 | 0,004 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,002 | 0,000 |
Scores machine (provisoires)
Les deux têtes enseignantes du modèle étudiant, lues sur ce travail. Un score ordonne la base pour la relecture; il n'affirme jamais une catégorie, et le statut de validation accompagne chaque rangée tel quel.
Scores de référence d'un modèle non mature (critères de maturité non atteints, 7 itérations). Un score ordonne; il n'affirme jamais une catégorie.
score_only:v0-immature-baseline · tel quel depuis la passe de notation : score_only signifie que le nombre peut ordonner les travaux, et qu'aucune étiquette de catégorie n'en découle