Preparation of true solutions of monomeric amyloidogenic protein/peptide：A critical prerequisite for aggregation kinetic study

• Published in: Science China. Chemistry Vol. 55; no. 1; pp. 118 - 124
• Main Authors: Diao, Shu, Zhao, Hong, Wang, WeiMao, Wu, Chi
• Format: Journal Article
• Language: English
• Published: Heidelberg SP Science China Press 2012; Springer Nature B.V
• Subjects: Aggregates; Chemistry; Chemistry and Materials Science; Chemistry/Food Science; Dissolution; Kinetics; Peptides; Proteins; Size distribution; 关联动力学; 制备; 动态激光光散射; 单体; 多肽; 正解; 淀粉样蛋白; 聚合动力学; laser light scattering; amyloidogenic protein/peptide; monomer; ultra-filtration
• ISSN: 1674-7291; 1869-1870
• DOI: 10.1007/s11426-011-4446-0

Our dynamic laser light scattering（LLS） study shows that the current widely used protocols of dissolving amyloidogenic protein/peptide do not really result in a true solution;namely,there always exist a trace amount of interchain aggregates,which greatly affect the association kinetics,partially explaining why different kinetics were reported even for a solution with identical protein and solvent.Recently,using a combination of the conventional dissolution procedure and our newly developed ultra-filtration method,we have developed a novel protocol to prepare a true solution of amyloidogenic protein/peptide without any interchain aggregates.The resultant solutions remain in their monomeric state for at least one week,which is vitally important for further study of the very initial stage of the interchain association under the physiological conditions because more and more evidence suggests that it is those small oligomers rather than large fabric aggregates that are cytotoxic.In addition,this study shows that combining static and dynamic LLS can lead to more physical and microscopic information about the protein association instead of only the size distribution.