title: Probing protein surface accessibility of amino acid substitutions using hydrophobic interaction chromatography.
abstract: Hydrophobic interaction chromatography (HIC) has been used to determine the influence of amino acid substitutions on protein retention and thereby their accessibility on the protein surface. The retentions of mutants of green fluorescent protein (GFPuv) and human hemoglobin (Hb) were studied on multimodal HIC media and compared to the hydrophobicities from known hydrophobicity scales with respect to the accessible surface area. For GFPuv, the theoretical and experimental results of three hydrophobicity scales correlated well (R(2)>0.85), which clearly indicate that the results can be used for protein retention prediction as well as probing surface properties of protein variants.
authors.affiliation: Department of Pure and Applied Biochemistry, Center for Chemistry and Chemical Engineering, Lund University, P.O. Box 124, SE-211 00 Lund, Sweden.
authors.forename: Kristian
authors.initials: K
authors.lastname: Becker
authors.forename: Marie
authors.initials: M
authors.lastname: Grey
authors.forename: Leif
authors.initials: L
authors.lastname: Bülow
mesh_terms: D019943:Amino Acid Substitution; D002853:Chromatography, Liquid / Q000379:methods*; D008958:Models, Molecular; D011506:Proteins / Q000737:chemistry*; D013499:Surface Properties
chemical_list: D011506:Proteins