Time-course, negative-stain electron microscopy-based analysis for investigating protein-protein interactions at the single-molecule level.

ABSTRACT: Several biophysical approaches are available to study protein-protein interactions. Most approaches are conducted in bulk solution, and are therefore limited to an average measurement of the ensemble of molecular interactions. Here, we show how single-particle EM can enrich our understanding of protein-protein interactions at the single-molecule level and potentially capture states that are unobservable with ensemble methods because they are below the limit of detection or not conducted on an appropriate time scale. Using the HIV-1 envelope glycoprotein (Env) and its interaction with receptor CD4-binding site neutralizing antibodies as a model system, we both corroborate ensemble kinetics-derived parameters and demonstrate how time-course EM can further dissect stoichiometric states of com

SUBMITTER: Nogal B 

PROVIDER: S-EPMC5702678 | biostudies-literature | 2017 Nov

REPOSITORIES: biostudies-literature