title: Comparisons of the structures of the chromosomal high mobility group proteins HMG1 and HMG2 prepared under conditions of neutral and acidic pH.
abstract: The chromosomal proteins HMG1 and 2 have been prepared by salt extraction and phosphocellulose chromatography at neutral pH (Isackson, P.J., Debold, W.A. and Reeck, G.R. (1980) FEBS Lett. 119, 337-342) to minimize protein denaturation. The structures of these phosphocellulose-prepared high mobility group proteins have been compared with those of high mobility group proteins using the previously described acid-extraction conditions which fully denature the proteins. When compared in the same solvent conditions the acid-extracted proteins did not refold to give the same level of alpha-helical and tertiary folded structures as the phosphocellulose-prepared proteins, suggesting that acid treatment can cause some irreversible damage to the proteins. This finding was supported by changes in the structure observed when phosphocellulose-prepared HMG1 was neutralized after exposure to acid. Gel filtration studies reveal no differences in the size of the high mobility group proteins, phosphocellulose-prepared and acid-extracted proteins both being largely monomeric in solution. Little difference was detected in the DNA-binding properties of the two types of protein, nor was there any difference in the oxidation state of the cysteines. However, isoelectric focusing analysis revealed differences in the subfractions of HMG2 prepared by the two methods.
mesh_terms: D000818:Animals; D002850:Chromatography, Gel; D002868:Chromosomal Proteins, Non-Histone / Q000302:isolation & purification*; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D006609:High Mobility Group Proteins; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D009682:Magnetic Resonance Spectroscopy; D011487:Protein Conformation; D013552:Swine
chemical_list: D002868:Chromosomal Proteins, Non-Histone; D006609:High Mobility Group Proteins