Purification of Casein Kinase II From Lung Tissue  and Kinetic Evaluations With Polyamines and  Myc-Oncoprotein

Nesrin GÜNDOĞUŞ-ÖZCANLI

Makale detayları

Purification of Casein Kinase II From Lung Tissue and Kinetic Evaluations With Polyamines and Myc-Oncoprotein

Yazarlar:

Nesrin GÜNDOĞUŞ-ÖZCANLI

Yayıncı:

TUBITAK

Dergi:

Turkish Journal of Medical Sciences 1998 Vol.28 Issue 6, pp.599-604

Belge Tipi:

Makale

Yayın Tarihi:

5/1998

Belge Dili:

Türkçe

Document ID (Org No.):

2000758165

Sıklık:

İki Aylık

eISSN:

1303-6165

pISSN:

1300-0144

Anahtar kelimeler:

Özet:

 

The overall goal of this study was  to seek molecular linkages between  polyamines, casein kinase II, and the myc  oncoprotein by using a cell-free, highly  purified enzyme kinetic analysis. Casein  kinase II (CKII) was purified from sheep lung  tissue by using an original 3-column/3-day  set of procedures (DEAE cellulose, Sepharose  6L-4B, and poly-L- lysine agrose affinity  chromotographies). CKII was purified  approximately 550-fold with a recovery of  near 40% and a final specific activity of about  250,000 pmol of phosphorylated protein  substrate (casein)/min/mg of enzyme protein.  Kinetic evaluations with the purified CKII  using Mg-GTP as a phosphate donor, etiher  casein or myc oncoprotein as substrate, and  various polyamines (polylysine, polyornithine,  spermidine, or spermine) as stimulators  showed: 1) phosphorylation of casein  icreased 2-4 fold with various polyamines; 2)  without polyamines, phosphorylation of myc  oncoprotein was about 10-20% when  compared to casein; 3) phosphorylation of  myc oncoprotein increased 10- to 30-fold in  the presence of various polyamines. The  phosphorylated form of myc oncoprotein is  the only biologically active form which  transactivates specific genes related to cell  proliferation.

 

The overall goal of this study was  to seek molecular linkages between  polyamines, casein kinase II, and the myc  oncoprotein by using a cell-free, highly  purified enzyme kinetic analysis. Casein  kinase II (CKII) was purified from sheep lung  tissue by using an original 3-column/3-day  set of procedures (DEAE cellulose, Sepharose  6L-4B, and poly-L- lysine agrose affinity  chromotographies). CKII was purified  approximately 550-fold with a recovery of  near 40% and a final specific activity of about  250,000 pmol of phosphorylated protein  substrate (casein)/min/mg of enzyme protein.  Kinetic evaluations with the purified CKII  using Mg-GTP as a phosphate donor, etiher  casein or myc oncoprotein as substrate, and  various polyamines (polylysine, polyornithine,  spermidine, or spermine) as stimulators  showed: 1) phosphorylation of casein  icreased 2-4 fold with various polyamines; 2)  without polyamines, phosphorylation of myc  oncoprotein was about 10-20% when  compared to casein; 3) phosphorylation of  myc oncoprotein increased 10- to 30-fold in  the presence of various polyamines. The  phosphorylated form of myc oncoprotein is  the only biologically active form which  transactivates specific genes related to cell  proliferation.

Makale detayları

Purification of Casein Kinase II From Lung Tissue and Kinetic Evaluations With Polyamines and Myc-Oncoprotein

Yazarlar:

Nesrin GÜNDOĞUŞ-ÖZCANLI

Yayıncı:

TUBITAK

Dergi:

Turkish Journal of Medical Sciences 1998 Vol.28 Issue 6, pp.599-604

Belge Tipi:

Makale

Yayın Tarihi:

5/1998

Belge Dili:

Türkçe

Document ID (Org No.):

2000758165

Sıklık:

İki Aylık

eISSN:

1303-6165

pISSN:

1300-0144

Casein kinase II

polyamines

myc

cell proliferation abbreviations: CKII-casein kinase II; Pmyc-phosphorylated myc oncoprotein; Pmax-phosphorylated max protein.

Sayı

Makale detayları

Purification of Casein Kinase II From Lung Tissue and Kinetic Evaluations With Polyamines and Myc-Oncoprotein

Yazarlar: Nesrin GÜNDOĞUŞ-ÖZCANLI

Yayıncı: TUBITAK

Dergi: Turkish Journal of Medical Sciences 1998 Vol.28 Issue 6, pp.599-604

Belge Tipi: Makale

Yayın Tarihi: 5/1998

Belge Dili: Türkçe

Document ID (Org No.): 2000758165

Sıklık: İki Aylık

eISSN: 1303-6165

pISSN: 1300-0144

Casein kinase II

polyamines

myc

cell proliferation abbreviations: CKII-casein kinase II; Pmyc-phosphorylated myc oncoprotein; Pmax-phosphorylated max protein.

Sayı

Yazarlar:

Nesrin GÜNDOĞUŞ-ÖZCANLI

Yayıncı:

TUBITAK

Dergi:

Turkish Journal of Medical Sciences 1998 Vol.28 Issue 6, pp.599-604

Belge Tipi:

Makale

Yayın Tarihi:

5/1998

Belge Dili:

Türkçe

Document ID (Org No.):

2000758165

Sıklık:

İki Aylık

eISSN:

1303-6165

pISSN:

1300-0144