Premium
Expanding the scope of CE reactor to ssDNA‐binding protein–ssDNA complexes as exemplified for a tool for direct measurement of dissociation kinetics of biomolecular complexes
Author(s) -
Takahashi Toru,
Ohtsuka Keiichirou,
Tomiya Yoriyuki,
Iki Nobuhiko,
Hoshino Hitoshi
Publication year - 2009
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200900110
Subject(s) - dissociation (chemistry) , kinetics , chemistry , kinetic energy , analytical chemistry (journal) , chromatography , physics , quantum mechanics
CE reactor (CER), which was developed as a tool for direct measurement of the dissociation kinetics of metal complexes, was successfully applied to the complexes of Escherichia coli ssDNA‐binding protein (SSB) with ssDNA. The basic concept of CER is the application of CE separation process as a dissociation kinetic reactor for the complex, and the observation of the on‐capillary dissociation reaction profile of the complex as the decrease of the peak height of the complex with increase of the migration time. The peak height of [SSB‐ssDNA] decreases as the migration time increases since the degree of the decrease of [SSB‐ssDNA] through the on‐capillary dissociation reaction is proportional to the degree of the decrease of the peak height of [SSB‐ssDNA]. The dissociation degree‐time profiles for the complexes are quantitatively described by analyzing a set of electropherograms with different migration times. Dissociation rate constants of [SSB‐ssDNA] consisting of 20‐mer, 25‐mer and 31‐mer ssDNA were directly determined to be 3.99×10 −4 , 4.82×10 −4 and 1.50×10 −3 /s, respectively. CER is a concise and effective tool for dissociation kinetic analysis of biomolecular complexes.