Aggregation and Oligomerization Characterization of ß-Lactoglobulin Protein Using a Solid-State Nanopore Sensor

Authors

Mitu C. Acharjee, University of Arkansas, FayettevilleFollow
Brad Ledden, University of Arkansas, Fayetteville
Brian Thomas, University of Arkansas, Fayetteville
Xianglan He, Rutgers UniversityFollow
Troy Messina, Berea College
Jason Giurleo, Regeneron PharmaceuticalsFollow
David Talaga, Montclair State University
Jiali Li, University of Arkansas, FayettevilleFollow

Document Type

Article

Publication Date

1-2024

Keywords

protein aggregation; solid-state nanopore; ss-lactoglobulin; protein oligomerization; protein volume

Abstract

Protein aggregation is linked to many chronic and devastating neurodegenerative human diseases and is strongly associated with aging. This work demonstrates that protein aggregation and oligomerization can be evaluated by a solid-state nanopore method at the single molecule level. A silicon nitride nanopore sensor was used to characterize both the amyloidogenic and native-state oligomerization of a model protein ß-lactoglobulin variant A (βLGa). The findings from the nanopore measurements are validated against atomic force microscopy (AFM) and dynamic light scattering (DLS) data, comparing βLGa aggregation from the same samples at various stages. By calibrating with linear and circular dsDNA, this study estimates the amyloid fibrils’ length and diameter, the quantity of the βLGa aggregates, and their distribution. The nanopore results align with the DLS and AFM data and offer additional insight at the level of individual protein molecular assemblies. As a further demonstration of the nanopore technique, βLGa self-association and aggregation at pH 4.6 as a function of temperature were measured at high (2 M KCl) and low (0.1 M KCl) ionic strength. This research highlights the advantages and limitations of using solid-state nanopore methods for analyzing protein aggregation.

Citation

Acharjee, M. C., Ledden, B., Thomas, B., He, X., Messina, T., Giurleo, J., Talaga, D., & Li, J. (2024). Aggregation and Oligomerization Characterization of ß-Lactoglobulin Protein Using a Solid-State Nanopore Sensor. Sensors, 24 (1), 81. https://doi.org/10.3390/s24010081

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.