Labeling of a Mutant Estrogen Receptor with an Affimer in a Breast Cancer Cell Line. Journal Article

Local Library Link: Find It @ Loyola

Authors: Ren, P; Tiede, C; Fanning, SW; Adams, T; Speirs, V; Nelson, ER; Cheng, C; Moore, TW; Greene, GL; Tomlinson, D; Selvin, PR
Article Title: Labeling of a Mutant Estrogen Receptor with an Affimer in a Breast Cancer Cell Line.
Abstract: Mutations of the intracellular Estrogen Receptor alpha (ERa) is implicated in 70% of breast cancers. It is therefore of considerable interest to image various mutants (L536S, Y537S, D538G) in living cancer cell lines, particularly as a function of various anti-cancer drugs. We therefore developed a small (13kD) Affimer, which, after fluorescent labeling, is able to efficiently label ERa by traveling through temporary pores in the cell membrane, created by the toxin Streptolysin O. The Affimer, selected by a phage display, predominantly labels the Y537S mutant and can tell the difference between L536S and D538G mutants. The vast majority of Affimer-ERaY537S is in the nucleus and is capable of an efficient, unrestricted navigation to its target DNA sequence, as visualized by single molecule fluorescence. The Affimer can also differentiate the effect of selective estrogen receptor modulators (SERMs). More generally, this is an example of a small binding reagent-an Affimer protein-that can be inserted into living cells with minimal perturbation and high efficiency, to image an endogenous protein.
Journal Title: Biophysical journal
ISSN: 1542-0086; 0006-3495
Publisher: Unknown  
Date Published: 2022