Abstract
The control of protein solubility is a subject of broad interest. 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 now present a method for screening solubilizing compounds, using an array of N- or C-terminal deletion mutants of the protein. A key behind this approach is that such terminal deletions of the protein affect its aggregation propensity. The solubilization activities of trial solvents are individually assessed, based on the number of solubilized mutants. The solubilizing compounds are then identified from the screened solvents. 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 s...Continue Reading
References
Nov 1, 1995·Journal of Biomolecular NMR·F DelaglioA Bax
Nov 14, 1997·Structure·N E Chayen
Dec 9, 1997·Journal of Biomolecular NMR·S BagbyM Ikura
Dec 23, 1998·Journal of Biomolecular NMR·C A Lepre, J M Moore
Feb 19, 2000·Protein Engineering·G LindwallL A Kohlstaedt
Jul 21, 2001·Methods in Enzymology·S BagbyM Ikura
Aug 3, 2001·The EMBO Journal·T ItoH Sumimoto
Aug 3, 2001·The EMBO Journal·H TerasawaF Inagaki
Oct 1, 2003·The EMBO Journal·Sosuke YoshinagaFuyuhiko Inagaki
Oct 2, 2004·Nature Medicine·Mark S FormanVirginia M-Y Lee
Mar 10, 2005·Journal of Biomolecular NMR·Peter W A Howe
Jun 3, 2005·Acta Crystallographica. Section D, Biological Crystallography·Thomas S WalterKarl Harlos
Jul 5, 2005·Nature Immunology·Yuya TerashimaKouji Matsushima
Sep 27, 2006·Acta Crystallographica. Section D, Biological Crystallography·Ian M BerryAlbrecht Messerschmidt
Nov 16, 2007·Proteins·Heath E KlockScott A Lesley
Jan 4, 2008·Protein Expression and Purification·Susanne GräslundJohan Weigelt
Feb 1, 2008·Nature Methods·UNKNOWN Structural Genomics ConsortiumKristin C Gunsalus
Apr 24, 2009·Protein Science : a Publication of the Protein Society·Toshihiko SugikiHideo Takahashi
Oct 21, 2009·The Journal of Immunology : Official Journal of the American Association of Immunologists·Etsuko TodaKouji Matsushima
Mar 2, 2010·International Journal of Pharmaceutics·Wei WangDirk Teagarden
Jul 14, 2010·Progress in Nuclear Magnetic Resonance Spectroscopy·Michael R GrykMark W Maciejewski
Jan 5, 2011·Protein Expression and Purification·Kaori EsakiHiroaki Terasawa
Mar 31, 2012·Trends in Pharmacological Sciences·Christopher W MurrayDavid C Rees
Oct 17, 2013·The Biochemical Journal·Etsuko TodaKouji Matsushima
Oct 7, 2014·The FEBS Journal·Kaori EsakiHiroaki Terasawa
Mar 28, 2017·Molecular Biotechnology·Akihiro SonodaHiroaki Terasawa