title: Proper calibration of ultrasonic power enabled the quantitative analysis of the ultrasonication-induced amyloid formation process.
abstract: To elucidate the mechanisms of ultrasonication on the amyloid fibril formation, we quantitatively determined the ultrasonic power using both calorimetry and potassium iodide (KI) oxidation, and under the properly calibrated ultrasonic power, we investigated the ultasonication-induced amyloid formation process of the mouse prion protein (mPrP(23-231)). These methods revealed that the ultrasonic power in our system ranged from 0.3 to 2.7 W but entirely dependent on the positions of the ultrasonic stage. Intriguingly, the nucleation time of the amyloid fibrils was found to be shortened almost proportionally to the ultrasonic power, indicating that the probability of the occurrence of nucleus formation increases proportionally to the ultrasonic power. Moreover, mPrP(23-231) formed two types of aggregates: rigid fibrils and short fibrils with disordered aggregates, depending on the ultrasonic power. The nucleation of rigid fibrils required an ultrasonic power larger than 1.5 W. While at the strong ultrasonic power larger than 2.6 W, amyloid fibrils were formed early, but simultaneously fine fragmentation of fibrils occurred. Thus, an ultrasonic power of approximately 2.0 W would be suitable for the formation of rigid mPrP(23-231) fibrils under the conditions utilized (ultrasonication applied for 30 s every 9 min). As ultrasonication has been widely used to amplify the scrapie form of the prion protein, or other amyloids in vitro, the calorimetry and KI oxidation methods proposed here might help determining the adequate ultrasonic powers necessary to amplify them efficiently.
authors.affiliation: Center for Emerging Infectious Diseases, Gifu University, Yanagido 1-1, Gifu, Japan.
authors.forename: Kei-ichi
authors.initials: K
authors.lastname: Yamaguchi
authors.forename: Tomoharu
authors.initials: T
authors.lastname: Matsumoto
authors.forename: Kazuo
authors.initials: K
authors.lastname: Kuwata
mesh_terms: D000682:Amyloid / Q000737:chemistry* / Q000378:metabolism*; D000818:Animals; D002138:Calibration; D002151:Calorimetry; D002942:Circular Dichroism; D019286:Endopeptidase K / Q000737:chemistry; D051379:Mice; D008854:Microscopy, Electron; D010084:Oxidation-Reduction; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D011193:Potassium Iodide / Q000737:chemistry; D011328:Prions / Q000737:chemistry / Q000378:metabolism; D013696:Temperature; D014465:Ultrasonics / Q000379:methods*
chemical_list: D000682:Amyloid; D010446:Peptide Fragments; D011328:Prions; C428437:prion protein (23-231); D011193:Potassium Iodide; D019286:Endopeptidase K
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