Use of fluorescence-detected sedimentation velocity to study high-affinity protein interactions.

ABSTRACT: Sedimentation velocity (SV) analytical ultracentrifugation (AUC) is a classic technique for the real-time observation of free macromolecular migration in solution driven by centrifugal force. This enables the analysis of macromolecular mass, shape, size distribution, and interactions. Although traditionally limited to determination of the sedimentation coefficient and binding affinity of proteins in the micromolar range, the implementation of modern detection and data analysis techniques has resulted in marked improvements in detection sensitivity and size resolution during the past decades. Fluorescence optical detection now permits the detection of recombinant proteins with fluorescence excitation at 488 or 561 nm at low picomolar concentrations, allowing for the study of high-affinity p

SUBMITTER: Chaturvedi SK 

PROVIDER: S-EPMC7466938 | biostudies-literature | 2017 Sep

REPOSITORIES: biostudies-literature