Probing protein surface accessibility of amino acid substitutions using hydrophobic interaction chromatography

Kristian Becker; Marie Grey; Leif Bülow

doi:10.1016/j.chroma.2008.11.002

Probing protein surface accessibility of amino acid substitutions using hydrophobic interaction chromatography

J Chromatogr A. 2008 Dec 26;1215(1-2):152-5. doi: 10.1016/j.chroma.2008.11.002. Epub 2008 Nov 7.

Authors

Kristian Becker¹, Marie Grey, Leif Bülow

Affiliation

¹ Department of Pure and Applied Biochemistry, Center for Chemistry and Chemical Engineering, Lund University, P.O. Box 124, SE-211 00 Lund, Sweden.

PMID: 19022452
DOI: 10.1016/j.chroma.2008.11.002

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Substitution*
Chromatography, Liquid / methods*
Models, Molecular
Proteins / chemistry*
Surface Properties

Substances

Proteins