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KMID : 0578319930030030275
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Molecules and Cells
1993 Volume.3 No. 3 p.275 ~ p.282
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Purification and Characterization of Histone H1 Kinase from Rat Liver
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Park, Iha
Oh, Sangtaek/Hong, Jeongho/Lee, Seung-Ki/Yim, Jeongbin
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Abstract
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A kinase which catalyzes the phosphorylation of histone H I was puri fied approximately 360 fold with a yield of 84%a from the normal rat liver. At first, we tried to purify and characterize the cdc2 kinase which regulates the entry into M phase of eukaryotic cells by the use of its histone H1 kinase activity. But the substrate specificity and other characteristics of the purified histone H1 kinase activity were found to be different from those of cdc2 kinase. In addition, Western blot analysis showed that the partially purified kinase appeared not to contain the cdc2 molecule. These results along with the phosphopeptide analysis indicate that histone HI kinase activity from normal liver was not due to cdc2 kinase. Molecular weight of the partially purified enzyme was estimated to be 37 kDa. The optimum temperature for the assay was 37 C and optimum pH was pH 7. The kinase activity was stable at 30 C for 30 min but lost the activity completely when treated above 50 T. Phosphate ion and Q-glycerophosphate inhibit the kinase activity. The Km´ value for ATP was 26.5 pM and Km for histone H1 was 0.74 pM. Among various cations tested, Sn2´ and Sn+ activated the kinase activity (15-20 fold), whereas divalent cations such as Cil", Ni", Zn` inhibited the kinase activity.
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