Constructing a Single Tryptophan Arrestin-3 Variant A3V9W by site-direct mutagenesis

Abstract

To study the potential binding-induced conformational changes on arrestin-3, a multifunctional adaptor protein, we intend to incorporate a fluorinated tryptophan residue as reporter in this protein, then obtain the fluorine NMR spectroscopy of this protein. A3V9W variant was designed to place a single tryptophan residue at the N-terminal of arrestin-3, which was reported as a primary binding site for some arrestin-3 binding partners such as JNK3 and ERK1/2. Here, we report our efforts to construct this arrestin-3 variant expression plasmid (pTrc HisB) by site-direct mutagenesis.