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Large-Scale Experimental Studies Show Unexpected Amino Acid Effects On Protein Expression And Solubility In Vivo In E. Coli, W. Nicholson Price Ii, Samuel K. Handelman, John K. Everett, Saichiu N. Tong, Ana Bracic, Jon D. Luff, Victor Naumov, Thomas Acton, Philip Manor, Rong Xiao, Burkhard Rost, Gaetano T. Montelione, John F. Hunt

Articles

The biochemical and physical factors controlling protein expression level and solubility in vivo remain incompletely characterized. To gain insight into the primary sequence features influencing these outcomes, we performed statistical analyses of results from the high-throughput protein-production pipeline of the Northeast Structural Genomics Consortium. Proteins expressed in E. coli and consistently purified were scored independently for expression and solubility levels. These parameters nonetheless show a very strong positive correlation. We used logistic regressions to determine whether they are systematically influenced by fractional amino acid composition or several bulk sequence parameters including hydrophobicity, sidechain entropy, electrostatic charge, and predicted backbone disorder. …