THE USE OF AFFINITY COELECTROPHORESIS TO CHARACTERIZE COOPERATIVE, NONSPECIFIC NUCLEIC-ACID BINDING PEPTIDES

被引:6
作者
NEDVED, ML [1 ]
MOE, GR [1 ]
机构
[1] UNIV DELAWARE,DEPT CHEM & BIOCHEM,NEWARK,DE 19716
关键词
D O I
10.1006/abio.1995.1255
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Affinity coelectrophoresis (ACE) is a technique for characterizing ligand/nucleic acid binding interactions under equilibrium conditions. It is used here to characterize the protamine clupeine Z binding to several DNA fragments in order to define the use and limitations of ACE for ligands that bind cooperatively and nonspecifically to nucleic acids. The results demonstrate that the ACE data for cooperative, nonspecific ligands can be analyzed using the McGhee-von Hippel model and that binding-site sizes can be accurately determined using lattices containing as few as one site. However, binding constants can be greatly underestimated for some cooperative ligands with large-site sizes if small lattices are used. The salt dependence of the binding constant can also be determined but is limited to salt concentrations less than similar to 300 mM. Given the simplicity and reproducibility of the ACE assay, it should find many applications for studying binding interactions for a variety of cooperative and noncooperative nucleic acid binding peptides and proteins. (C) 1995 Academic Press, Inc.
引用
收藏
页码:80 / 84
页数:5
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