Abstract
Peroxiredoxins (PRDX) are antioxidant enzymes that are critical to cellular function. Altered activity of these enzymes has been implicated in the development of neurodegenerative diseases, cancer, and inflammatory conditions. PRDXs exist in two conformations, a dimeric structure and a decameric structure. No methods are available to study the relationship between these two conformational states of PRDX and their activity in cells. Nanobodies (NB) are a new tool in the study of protein structure and function. NB consist of a single variable domain that has been derived from camelid antibodies, which contain only heavy chains. NB can bind to their target with high affinity and are conformation specific. Fluorescently tagged NB have been successfully used to study protein endogenously. The aim of this research was to generate fluorescently tagged PRDX specific nanobodies that could be used to study PRDX within the crowded cellular environment.