6533b823fe1ef96bd127eb98
RESEARCH PRODUCT
Fluctuation Methods To Study Protein Aggregation in Live Cells: Concanavalin A Oligomers Formation
Valeria MilitelloGiulia OssatoMaurizio LeoneMichelle A. DigmanValeria VetriEnrico Grattonsubject
Time FactorsCell SurvivalCellSpectroscopy Imaging and Other TechniquesBiophysicsProtein aggregationCell morphologyCell membraneDiffusion03 medical and health scienceschemistry.chemical_compoundMice0302 clinical medicineProtein structure2-NaphthylaminemedicineConcanavalin AAnimalsconfocal microscopy super resolution protein aggregation kinetics in live cells amyloid related pathologiesAnnexin A5Protein Structure QuaternaryCell Shape030304 developmental biology0303 health sciencesbiologySpectrum AnalysisCell MembraneFibroblastsEmbryo MammalianCell biologyMembranemedicine.anatomical_structurechemistryConcanavalin Abiology.proteinLaurdan030217 neurology & neurosurgeryFluorescein-5-isothiocyanateLauratesdescription
Prefibrillar oligomers of proteins are suspected to be the primary pathogenic agents in several neurodegenerative diseases. A key approach for elucidating the pathogenic mechanisms is to probe the existence of oligomers directly in living cells. In this work, we were able to monitor the process of aggregation of Concanavalin A in live cells. We used number and brightness analysis, two-color cross number and brightness analysis, and Raster image correlation spectroscopy to obtain the number of molecules, aggregation state, and diffusion coefficient as a function of time and cell location. We observed that binding of Concanavalin A to the membrane and the formation of small aggregates paralleled cell morphology changes, indicating progressive cell compaction and death. Upon protein aggregation, we observed increased membrane water penetration as reported by Laurdan generalized polarization imaging. © 2011 by the Biophysical Society.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2011-02-01 |