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Published on: October 23, 2018
Isolation and Partial Purification of Two Types of Nuclear Tyrosine Protein Kinase from Rat Liver
1Key Laboratory of Glycoconjugate Research, Ministry of Public Health, Department of Biochemistry, Shanghai Medical University, Shanghai 200032, China.
Abstract:
Nuclear tyrosine protein kinase (n-TPK) plays an important role in the regulation of cell cycle and gene transcription. In the present investigation, two types of n-TPK isozyme, n-TPK I and n-TPK II were isolated from the nuclear extract of rat liver by using DEAE-Sepharose CL-6B ion-exchange column chromatography. The two isozymes were further partially purified on heparin affinity column and Sephadex G-100 to 30 and 25 folds respectively. The specific activities of n-TPK I and n-TPK II were 1 638 and 1 360 pmol/(min.mg protein) respectively. By means of Sephadex G-100 chromatography and autoradiography of the autophosphorylated n-TPK after [32P]-ATP labeling and SDS-PAGE, the molecular weight of n-TPK I and n-TPK II were determined to be 76 kD and 59 kD respectively. Both n-TPK isozymes are monomers.
