Oleic acid inhibits amyloid formation of the intermediate of alpha-lactalbumin at moderately acidic pH.
The effects of oleic acid on amyloid formation of Ca2+-depleted bovine alpha-lactalbumin (apo-BLA) at low pH and the biological impact of the effects were investigated by using thioflavin T, Congo red, far-UV circular dichroism, atomic force microscopy, transmission electron microscopy, and other biophysical methods. The results from the phase diagram method of fluorescence show that two intermediates exist in the conformational transition of apo-BLA induced by low pH. One intermediate populated at pH 3.0 is characterized as a molten globule state and the other accumulates with stable secondary structure and exposed hydrophobic surface at pH 4.0-4.5. Amyloid formation of apo-BLA takes place upon decreasing the pH to 4.5 and is accelerated remarkably as the pH is decreased further. However, amyloid fibrils of apo-BLA are not observed in the pH range of 5.0-7.0 on a time-scale of 30 days. The lag time of fibrillation at pH 4.0 is greatly elongated by the presence of oleic acid, accompanied by a remarkable decline of the maximum thioflavin T intensity. Furthermore, amyloid formation of apo-BLA at pH 4.5 is inhibited completely by oleic acid, and insoluble aggregates are observed. In contrast, the effects of oleic acid on amyloid formation are not remarkable at pH 3.0 or at pH 2.0. Our data demonstrate that oleic acid specifically induces the intermediate of apo-BLA at pH 4.0-4.5 to form insoluble amorphous aggregates, which is responsible for the inhibition of amyloid formation of the protein by oleic acid in this range of pH values.
Yang
Fang
F
State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan 430072, China.
Zhang
Min
M
Zhou
Bing-Rui
BR
Chen
Jie
J
Liang
Yi
Y
D000682:Amyloid / Q000737:chemistry* / Q000648:ultrastructure; D000817:Anilino Naphthalenesulfonates / Q000378:metabolism; D000818:Animals; D001059:Apoproteins / Q000378:metabolism; D052160:Benzothiazoles; D002417:Cattle; D002942:Circular Dichroism; D003224:Congo Red / Q000378:metabolism; D005453:Fluorescence; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D007768:Lactalbumin / Q000737:chemistry* / Q000648:ultrastructure; D018625:Microscopy, Atomic Force; D046529:Microscopy, Electron, Transmission; D019301:Oleic Acid / Q000494:pharmacology*; D011485:Protein Binding / Q000187:drug effects; D020836:Protein Structure, Quaternary / Q000187:drug effects; D013056:Spectrophotometry, Ultraviolet; D013844:Thiazoles / Q000378:metabolism
D000682:Amyloid; D000817:Anilino Naphthalenesulfonates; D001059:Apoproteins; D052160:Benzothiazoles; D013844:Thiazoles; C009462:thioflavin T; D019301:Oleic Acid; D003224:Congo Red; D007768:Lactalbumin