Purification of histone deacetylase HD1-A of germinating maize embryos

FEBS Lett. 1996 Sep 16;393(2-3):287-91. doi: 10.1016/0014-5793(96)00909-x.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Nucleus / enzymology
  • Chromatography, Affinity
  • Chromatography, Gel
  • Chromatography, Ion Exchange
  • Electrophoresis, Polyacrylamide Gel
  • Histone Deacetylases / chemistry
  • Histone Deacetylases / isolation & purification*
  • Histone Deacetylases / metabolism
  • Hydrogen-Ion Concentration
  • Isoelectric Focusing
  • Kinetics
  • Molecular Weight
  • Seeds
  • Zea mays / enzymology*

Substances

  • Histone Deacetylases