Structural Analysis of the Effect of a Dual-FLAG Tag on Transthyretin.

ABSTRACT: Recombinant proteins have increased our knowledge regarding the physiological role of proteins; however, affinity purification tags are often not cleaved prior to analysis, and their effects on protein structure, stability and assembly are often overlooked. In this study, the stabilizing effects of an N-terminus dual-FLAG (FT₂) tag fusion to transthyretin (TTR), a construct used in previous studies, are investigated using native ion mobility-mass spectrometry (IM-MS). A combination of collision-induced unfolding and variable-temperature electrospray ionization is used to compare gas- and solution-phase stabilities of FT₂-TTR to wild-type and C-terminal tagged TTR. Despite an increased stability of both gas- and solution-phase FT₂-TTR, thermal degradation of

SUBMITTER: Shirzadeh M 

PROVIDER: S-EPMC7171973 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature