- 作者: 林淵泉
- 作者服務機構: Department of Chemistry, National Taiwan University, Taipei, Taiwan 107
- 中文摘要:
Using N,N′-Dicyclohexylcarbodiimide (DCC) as a condensing agent, 32P-cyanoethylphosphate and NH3 were combined to yield 32P-cyanoethylphosphoramidate. Treatment with NaOH resulted in a β-elimination step to yield 22P-phosphoramidate and acrylonitrile. The phosphoramidate was purified via DEAE-cellulose chromatography. At pH 7.4 the histidine residues of histone H4 were chemically phosphorylated using 32P-phosphoramidate as the phosphoryl donor. The nucleophilic nitrogen of the imidazole ring is attracted to the phosphorus of phosphoramidate resulting in the displacement of NH3 and formation of the protein phosphoryl-nitrogen linkages.
Chromatin monomers (nucleosomes) were prepared by a mild digestion of chromatin with micrococcal nuclease. This report also presents the chemical phosphorylation of these nucleosomes by 32P-phosphoramidate. - 英文摘要: --
- 中文關鍵字: --
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