Studying the Folding Pathway of Truncated and Full-Length Human Serum Albumin via FCS
Bunagan, Michelle R.
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Single molecule fluorescence correlation spectroscopy (FCS) can be used to determine the specific folding pathway of a fluorescently labeled protein by measuring the fluorescence intensity fluctuations caused by single molecules entering or leaving the observation volume. Here, Alexa-Fluor 488 labeled Human Serum Albumin (HSA) was characterized using FCS. This model protein was studied via FCS in a range of denaturant concentrations to identify occupied intermediate states along its folding pathway. A truncated HSA containing only Domain 1 was also studied. The FCS data was analyzed to extract the diffusion time associated with the protein molecules. The diffusion time can be used to calculate the radius of hydration of the labeled protein which is related to the protein conformation. Identifying the conformations of both the full-length HSA and Domain 1 in different denaturant conditions will allow us to gain a better understanding of the specific folding pathway undergone by HSA during denaturation.
Department of Chemistry
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