Purification and characterization of the protein kinase eEF-2 isolated from rat liver cells

• Autorzy: Gajko A. , Galasinski W. , Gindzienski A.
• Języki publikacji: EN

Abstrakty

The elongation factor 2 (eEF-2) protein kinase was isolated from rat liver cells, purified and partly characterized. It was found that the enzyme exists in an inactive form in the homogenate of rat liver. The active fraction of kinase eEF-2 was obtained after removal of the inhibitory substance by hydroxyapatite column chromatography. The purified enzyme is an electTophoretically homogeneous protein with relative molecular mass of approximately 90000 and isoelectric point, pi = 5.9. The enzyme specifically phosphorylates the elongation factor eEF-2 in the presence of calmodulin and Ca2+.

Słowa kluczowe

EN

cell; elongation factor; purification; protein; liver; rat; protein kinase; kinase

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 1994
• Tom: 41
• Numer: 4
• Opis fizyczny: p.421-427,fig.

Twórcy

autor

Gajko A.

• Medical Academy, 15-230 Bialystok 8, Poland

autor

Galasinski W.

autor

Gindzienski A.

Bibliografia

• 1. Nairn, A.C., Bhagat, B. & Palfrey, H.C. (1985) Identification of calmodulin-dependent protein kinase and its major Mr 100000 substrate in mammalian tissues. Prvc. Natl. Acad. Sci. U.S.A. 82,7939-7943.
• 2. Ryazanov, A.C., Natapov, P.G., Shestakova, E.A., Severin, F.F. & Spirin, A.S. (1988) Phosphorylation of the elongation factor 2: the fifth Ca2 /calmodulin dependent system of protein phosphorylation. Biochimie 70,619-626.
• 3. Nilsson, A., Carlberg, U. & Nygard, O. (1991) Kinetic characterization of the enzymatic activity of the EF-2 specific Ca2+- and calmo­dulin-dependent protein kinase III from rabbit reticulocytes. Eur. j. Biochem. 195,377-383.
• 4. Nygard, O., Nilsson, A., Carlberg, U., Nilsson, L. & Amons, R. (1991) Phosphorylation regulates the activity of the eEF-2 specific Ca2*- and calmodulin-dependent protein kinase III. /. Biol. Chan. 266,16425-16430.
• 5. Skogerson, L. &. Moldave, K. (1969) The binding of aminoacyltransferase II to ribosomes. Biochem. Biophys. Res. Commun. 27,568-572.
• 6. Galasiriski, W. & Moldave, K. (1969) Purification of aminoacyltransferase II (translocation factor) from rat liver. /. Biol. Chan. 244,6527-6532.
• 7. Moldave, K. (1963) Tine preparation of 14C-ami- noacyl soluble t-RNA. Methods Eiizymol. 6, 757-761.
• 8. Leyne, E. (1957) Spectrophotometric and turbidimcric methods for measuring proteins. Methods Lnzymol. 3,454.
• 9. Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochan. 72,248-254.
• 10. Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) 227,680-685.
• 11. Stahl, J., Welfle, H. & Bielka, H. (1972) Studies on protein of animal ribosomes. XIV. Analysis of phosphorylatcd rat liver ribosomal proteins by two dimensional polyacrylamide gel electro­phoresis. FEBS Lett. 26,233-236.
• 12. Gajko, A., Sredziriska, K., Marcinkiewicz, C. & Galasinski, W. (1991) The effect of phospho­rylation of the EF-2 isolated from rat liver cells on protein biosynthesis in vitro. Acta Biochim. Poloiu 38,353-358.
• 13. Tao, M. & Machet, P. (1973) Adenosine cyclic 3',5'-monophosphate-dependent protein kinase from rabbit reticulocytes. J. Biol. Chan. 24S, 5324-5332.
• 14. O'Far roll, P.H. (1975) High resolution two- -dimensional electrophoresis of proteins. J. Biol. Chan. 250,4007-4021.
• 15. Galasinski, W., Marcinkiewicz, C., Gajko, A., §redziriska, K. & 'Ielejko, E. (1993) Elongation factors from Guerin epithelioma and rat liver cells. Acta Biochim. Poton. 40,433-443.