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The dissociation rate of estrogen receptor alpha from the consensus estrogen response element.

Abstract：

:The rate of dissociation of recombinant, purified human estrogen receptor alpha (ERalpha) from a fluorescein-labeled DNA containing the consensus vitellogenin ERE sequence (F-vitERE) was determined in real time using fluorescence anisotropy. The complex of estradiol-occupied ERalpha with F-vitERE had an apparent dissociation rate of 1.48+/-0.06x10(-2) s(-1) and a half-life of 46.6 s at room temperature. The dissociation rate was characterized by a single exponential decay, suggesting that ER dissociates from the DNA as a preformed dimer, rather than as two individual monomers. The association rate of estradiol-occupied ERalpha for the F-vitERE was calculated as 7x10(6) M(-1) s(-1) based on the dissociation rate measured and previous determinations of the equilibrium dissociation constant (Kd) in similar assay conditions (Ozers et al., 1997). In buffer containing various concentrations of salt, the rate of dissociation of estradiol-occupied ERalpha from F-vitERE was accelerated by increasing salt concentrations. Compared to estradiol-occupied ERalpha, the rate of dissociation of unoccupied ERalpha from the F-vitERE was very similar, indicating that estradiol occupancy does not affect the dissociation rate of ERalpha from the ERE.

journal_name

Mol Cell Endocrinol

journal_title

Molecular and cellular endocrinology

authors

Szatkowski Ozers M,Hill JJ,Ervin K,Royer CA,Gorski J

doi

10.1016/s0303-7207(01)00395-1

keywords:

subject

Has Abstract

pub_date

2001-04-25 00:00:00

pages

101-9

issue

1-2

eissn

0303-7207

issn

1872-8057

pii

S0303-7207(01)00395-1

journal_volume

175

pub_type

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