In-cell 31P solid-state NMR measurements of the lipid dynamics and influence of exogeneous ß-amyloid peptides on live neuroblastoma neuro-2a cells.
Biophys Chem
; 297: 107008, 2023 06.
Article
in En
| MEDLINE
| ID: mdl-36989875
ABSTRACT
Non-specific disruption of cellular membranes induced by aggregation of exogeneous ß-amyloid (Aß) peptides is considered a viable pathological mechanism in Alzheimer's disease (AD). The solid-state nuclear magnetic resonance (ssNMR) spectroscopy has been widely applied in model liposomes to provide important insights on the molecular interactions between membranes and Aß aggregates. Yet, the feasibility of in-cell ssNMR spectroscopy to probe Aß-membrane interactions in native cellular environments has rarely been tested. Here we report the application of in-cell31P ssNMR spectroscopy on live mouse neuroblastoma Neuro-2a (N2a) cells under moderate magic angle spinning (MAS) conditions. Both cell viability and cytoplasmic membrane integrity are retained for up to six hours under 5 kHz MAS frequency at 277 K, which allow applications of direct-polarization 31P spectroscopy and 31P spin-spin (T2) relaxation measurements. The 31P T2 relaxation time constant of N2a cells is significantly increased compared with the model liposome prepared with comparable major phospholipid compositions. With the addition of 5 µM 40-residue Aß (Aß1-40) peptides, the 31P T2 relaxation is instantly accelerated. This work demonstrates the feasibility of using in-cell31P ssNMR to investigate the Aß-membrane interactions in the biologically relevant cellular system.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Alzheimer Disease
/
Neuroblastoma
Limits:
Animals
Language:
En
Journal:
Biophys Chem
Year:
2023
Type:
Article
Affiliation country:
United States