Efficient identification of compounds suppressing protein precipitation via solvent screening using serial deletion mutants of the target protein

In this study, the C‐terminal chemokine receptor‐binding region of the cytoplasmic protein, FROUNT (FNT‐C), which mediates intracellular signals leading to leukocyte migration, was subjected to the multicomponent screening. In total, 192 solution conditions were tested, using eight terminal deletion mutants of FNT‐C. We identified five solvent conditions that solubilized four or five mutants of FNT‐C, and the compounds in the screened solvents were then, respectively, assessed in terms of their solubilization ability. The best compound for solubilizing FNT‐C was 1,6‐hexanediol. Indeed, 1,6‐hexanediol bound to FNT‐C and suppressed its precipitation, as showed by NMR and dynamic light scattering analyses.

Although several solvent screening methods are available to search for compounds that enhance protein solubilization, their performance is influenced by the intrinsic solubility of the tested protein. We present a method for screening solubilizing compounds, using an array of N‐ or C‐terminal deletion mutants of the protein. Such terminal deletions of the protein affect its aggregation propensity.

Source: Genes to Cells - January 2, 2018 Category: Genetics & Stem Cells Authors: Kaori Yunoki, Sosuke Yoshinaga, Mitsuhiro Takeda, Ryohei Nagano, Yusuke Tsuchiya, Akihiro Sonoda, Tatsuichiro Tsuji, Makoto Hirakane, Etsuko Toda, Yuya Terashima, Kouji Matsushima, Hiroaki Terasawa Tags: ORIGINAL ARTICLE Source Type: research