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Histone kinase activity of buffalo sperm chromatin.

DIR@IMTECH: CSIR-Institute of Microbial Technology

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Title Histone kinase activity of buffalo sperm chromatin.
 
Creator Mudgal, P
Varshney, Grish C
Anand, S R
 
Subject QH426 Genetics
 
Description Phosphorylation of buffalo sperm chromatin proteins under optimum conditions (8 mM Mg2+, pH 8.0, and at 30 degrees C) using [gamma-32P]ATP and endogenous protein kinase activity was linear for 15 min incubation time and up to 330 micrograms protein. The 32P transferred from [gamma-32P]ATP was located in protein as a phosphoester bond. Fractionation with 1.2 M NaCl-4 M urea-0.2 M 2-mercaptoethanol-1 mM PMSF followed by acid treatment solubilized 87% of the total chromatin proteins termed "sperm histones." The remaining 21% nonhistone protein was tightly bound to DNA. Follow-up of the label showed 91% of the 32P in sperm histone and 9% with DNA-associated proteins. Histone kinase activity was solubilized with 0.35 M NaCl, which extracted 70% of the initial enzyme activity associated with chromatin. Of the different histones tested as substrates, histone kinase phosphorylated only histone H3 and, therefore, is highly specific for arginine rich histone. It also phosphorylates the acidic protein, casein. Cyclic AMP at concentrations up to 50 microM had no effect on the phosphorylation of histone H3. Phosphoamino acid analysis using histone H3 as the substrate showed serine to be the acceptor amino acid for phosphoester link.
 
Publisher Elsevier/North-Holland
 
Date 1997
 
Type Article
PeerReviewed
 
Identifier Mudgal, P and Varshney, Grish C and Anand, S R (1997) Histone kinase activity of buffalo sperm chromatin. Archives of andrology, 38 (3). pp. 191-9. ISSN 0148-5016
 
Relation http://crdd.osdd.net/open/768/