title: Mechanism of protein hydrophobic chromatography. Protein unfolding and its contribution to effective hydrophobicity.
abstract: Several different monomeric proteins with either one or two domains were used to study the effect of protein unfolding on effective hydrophobicity of proteins. Protein unfolding was inhibited by cross-linking with either toluene di-isocyanate or glutaraldehyde. The native enzyme was much more readily bound to the hydrophobic resin than the cross-linked species. The ligand, 3-phosphoglycerate, significantly decreased the amount of phosphoglycerate kinase able to bind to octyl-Sepharose. Sucrose also caused a statistically significant decrease in protein binding to the hydrophobic resin. These studies support the concept that the degree of protein unfolding influences effective hydrophobicity as measured by retention on octyl-Sepharose.
mesh_terms: D055598:Chemical Phenomena; D002621:Chemistry; D002850:Chromatography, Gel / Q000379:methods; D010735:Phosphoglycerate Kinase / Q000302:isolation & purification; D011487:Protein Conformation; D011489:Protein Denaturation; D011506:Proteins / Q000302:isolation & purification*; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D012685:Sepharose / Q000031:analogs & derivatives
chemical_list: D011506:Proteins; C027176:octyl-sepharose CL-4B; D012685:Sepharose; D010735:Phosphoglycerate Kinase