Isolation of high-mobility-group proteins HMG1 and HMG2 in non denaturing conditions and comparison of their properties with those of acid-extracted proteins.
We describe a method for isolation and purification of the chromosomal proteins HMG1 and HMG2 in non-denaturing conditions which overcomes the difficulties of the published methods concerning yield and purity. The method is based on salt extraction, selective precipitation with ammonium sulfate and DEAE-cellulose chromatography. All studied properties of these proteins (formation of protein tetramers, enhancement of micrococcal nuclease digestion of DNA and chromatin, and protection of 165-basepair DNA in chromatosome) differ significantly from the properties of HMG1 and 2 isolated under denaturing conditions.
D000818:Animals; D002417:Cattle; D002499:Centrifugation, Density Gradient; D002843:Chromatin / Q000378:metabolism; D002868:Chromosomal Proteins, Non-Histone / Q000302:isolation & purification*; D004247:DNA / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006609:High Mobility Group Proteins; D008722:Methods; D008836:Micrococcal Nuclease / Q000378:metabolism; D011489:Protein Denaturation
D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D006609:High Mobility Group Proteins; D004247:DNA; D008836:Micrococcal Nuclease