Biomaterial Database

Purification of rat liver nuclear protein kinase NII. 1977

W Thornburg, and T J Lindell

Rat liver nuclear protein kinase activity (NII), which is eluted from DEAE-Sephadex columns, has been purified approximately 1500-fold from solubilized nuclear protein. The method of purification involved chromatography of protein eluted from DEAE-Sephadex successively on phosvitin-Sepharose, mixed histone-Sepharose, and histone H2b-Sepharose followed by gel filtration on Sephadex G-200. Resulting preparations are homogeneous by polyacrylamide gel electrophoresis. The enzyme consists of three polypeptides with molecular weights of 42,000 (alpha), 39,000 (alpha'), and 26,000 (beta) which are present in the ratio 1:1:2 indicating that the enzyme has a minimum tetrameric subunit composition of alphaalpha'beta2. The molecular weight and s20,w of the purified enzyme were 123,000 and 7.0, respectively, as determined by sucrose density gradient centrifugation in 0.4 M NaCl. The enzyme has maximal activity with phosvitin as substrate and is not stimulated by 10(-5) to 10(-4) M cAMP or cGMP using H2b as substrate.

UI	MeSH Term	Description	Entries
D008099	Liver	A large lobed glandular organ in the abdomen of vertebrates that is responsible for detoxification, metabolism, synthesis and storage of various substances.	Livers
D008297	Male		Males
D008722	Methods	A series of steps taken in order to conduct research.	Techniques,Methodological Studies,Methodological Study,Procedures,Studies, Methodological,Study, Methodological,Method,Procedure,Technique
D010774	Phosvitin	An egg yolk phosphoglycoprotein which contains about 90% of the yolk protein phosphorus. It is synthesized in the liver of the hen and transferred to the developing oocyte, where it is bound to lipoproteins within the yolk granules.	Phosphovitellin
D011494	Protein Kinases	A family of enzymes that catalyze the conversion of ATP and a protein to ADP and a phosphoprotein.	Protein Kinase,Kinase, Protein,Kinases, Protein
D002467	Cell Nucleus	Within a eukaryotic cell, a membrane-limited body which contains chromosomes and one or more nucleoli (CELL NUCLEOLUS). The nuclear membrane consists of a double unit-type membrane which is perforated by a number of pores; the outermost membrane is continuous with the ENDOPLASMIC RETICULUM. A cell may contain more than one nucleus. (From Singleton & Sainsbury, Dictionary of Microbiology and Molecular Biology, 2d ed)	Cell Nuclei,Nuclei, Cell,Nucleus, Cell
D002846	Chromatography, Affinity	A chromatographic technique that utilizes the ability of biological molecules, often ANTIBODIES, to bind to certain ligands specifically and reversibly. It is used in protein biochemistry. (McGraw-Hill Dictionary of Scientific and Technical Terms, 4th ed)	Chromatography, Bioaffinity,Immunochromatography,Affinity Chromatography,Bioaffinity Chromatography
D004586	Electrophoresis	An electrochemical process in which macromolecules or colloidal particles with a net electric charge migrate in a solution under the influence of an electric current.	Electrophoreses
D006657	Histones	Small chromosomal proteins (approx 12-20 kD) possessing an open, unfolded structure and attached to the DNA in cell nuclei by ionic linkages. Classification into the various types (designated histone I, histone II, etc.) is based on the relative amounts of arginine and lysine in each.	Histone,Histone H1,Histone H1(s),Histone H2a,Histone H2b,Histone H3,Histone H3.3,Histone H4,Histone H5,Histone H7
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia