Purification of histone deacetylase HD1-A of germinating maize embryos.
We have purified the soluble nuclear histone deacetylase HD1-A of germinating maize embryos. By a combination of 6 chromatographic steps we achieved a 77,000-fold purification of an enzymatically active protein. Gel filtration chromatography revealed a molecular weight of 45 kDa of the native enzyme and electrophoretic analysis of the purified enzyme by SDS-PAGE resulted in a single band at a molecular weight of 48 kDa, indicating that the enzyme is a monomer protein. When fractions with enzyme activity of different stages of chromatographic purification were subjected to isoelectric focusing, enzyme activity focused at a pH of around 6.4 as measured in an activity gel assay; second dimension SDS-PAGE again revealed a protein spot at a molecular weight of 48 kDa.
Brosch
G
G
Department of Microbiology, University of Innsbruck, Medical School, Austria.
Goralik-Schramel
M
M
Loidl
P
P
D002467:Cell Nucleus / Q000201:enzymology; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D006655:Histone Deacetylases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D007700:Kinetics; D008970:Molecular Weight; D012639:Seeds; D003313:Zea mays / Q000201:enzymology*
D006655:Histone Deacetylases