Purification and characterization of p27, a protein from hepatocyte chromatin Evidence suggesting that it binds selectively to guanine‐rich single‐stranded DNA

FEBS Letters

Rahat, Michal A.¹  Fry, Michael¹ 

[1] Unit of Biochemistry, The Bruce Rappaport Faculty of Medicine, Technion - Israel Institute of Technology, POB 9649, Bat Galim, Haifa 31096, Israel

关键词: DNA binding protein;    Guanine-rich DNA;    Single-stranded DNA;    DE-52;    (diethylaminoethyl)cellulose-52;    DTT;    dithiothreitol;    HEPES;    (N[2-hydroxyethyl]piperazine-N′-[2-ethanesulfonic acid]);    P-11;    phosphocellulose 11;    MalNet;    N-ethylmaleimide;    PMSF;    phenylmethylsulfonylfluoride;   

DOI  :  10.1016/0014-5793(93)81680-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A protein designated p27 that binds preferentially to single-stranded DNA rich in guanine tracts was purified to near homogeneity from rabbit hepatocyte non-histone protein extract. Purified p27 migrated as a 27 kDa polypeptide on denaturing SDS-PAGE and displayed a native molecular mass of ∼ 155 kDa on Sephadex G-150 or Sepharose 6B-Cl gel filtration columns. Gel shift analysis indicated that maximum binding of p27 to single-stranded DNA required the presence of tracts of four or more contiguous guanine residues. The lowest found dissociation constant, 1.4 × 10⁻⁸ M/l, was for single-stranded DNA that contained a (dG)₁₇ run.

【 授权许可】

Unknown