A HaloTag Anchored Ruler for Week-Long Studies of Protein Dynamics.

ABSTRACT: Under physiological conditions, protein oxidation and misfolding occur with very low probability and on long times scales. Single-molecule techniques provide the ability to distinguish between properly folded and damaged proteins that are otherwise masked in ensemble measurements. However, at physiological conditions these rare events occur with a time constant of several hours, inaccessible to current single-molecule approaches. Here we present a magnetic-tweezers-based technique that allows, for the first time, the study of folding of single proteins during week-long experiments. This technique combines HaloTag anchoring, sub-micrometer positioning of magnets, and an active correction of the focal drift. Using this technique and protein L as a molecular template, we generate a magnet law

SUBMITTER: Popa I 

PROVIDER: S-EPMC5510598 | biostudies-literature | 2016 Aug

REPOSITORIES: biostudies-literature