Paredes Jose M, Casares Salvador, Ruedas-Rama Maria J, Fernandez Elena, Castello Fabio, Varela Lorena, Orte Angel
Department of Physical Chemistry, Faculty of Pharmacy, Campus Cartuja, Granada, 18071, Spain.
Department of Physical Chemistry, Faculty of Sciences, Campus Fuentenueva, Granada, 18071, Spain.
Int J Mol Sci. 2012;13(8):9400-9418. doi: 10.3390/ijms13089400. Epub 2012 Jul 25.
Amyloidogenic protein aggregation is a persistent biomedical problem. Despite active research in disease-related aggregation, the need for multidisciplinary approaches to the problem is evident. Recent advances in single-molecule fluorescence spectroscopy are valuable for examining heterogenic biomolecular systems. In this work, we have explored the initial stages of amyloidogenic aggregation by employing fluorescence lifetime correlation spectroscopy (FLCS), an advanced modification of conventional fluorescence correlation spectroscopy (FCS) that utilizes time-resolved information. FLCS provides size distributions and kinetics for the oligomer growth of the SH3 domain of α-spectrin, whose N47A mutant forms amyloid fibrils at pH 3.2 and 37 °C in the presence of salt. The combination of FCS with additional fluorescence lifetime information provides an exciting approach to focus on the initial aggregation stages, allowing a better understanding of the fibrillization process, by providing multidimensional information, valuable in combination with other conventional methodologies.
淀粉样蛋白聚集是一个长期存在的生物医学问题。尽管在与疾病相关的聚集方面进行了积极的研究，但显然需要多学科方法来解决这个问题。单分子荧光光谱的最新进展对于研究异质生物分子系统很有价值。在这项工作中，我们通过采用荧光寿命相关光谱（FLCS）探索了淀粉样蛋白聚集的初始阶段，FLCS是传统荧光相关光谱（FCS）的一种先进改进，它利用时间分辨信息。FLCS提供了α-血影蛋白SH3结构域寡聚体生长的尺寸分布和动力学信息，其N47A突变体在pH 3.2和37°C且存在盐的条件下形成淀粉样纤维。FCS与额外的荧光寿命信息相结合，为关注初始聚集阶段提供了一种令人兴奋的方法，通过提供多维信息，与其他传统方法相结合时很有价值，从而能更好地理解纤维化过程。
