Linked Life Data

7171576

Source:http://linkedlifedata.com/resource/pubmed/id/7171576

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
22
pubmed:dateCreated
1983-2-25
pubmed:abstractText
5'-Deoxypyridoxal, a vitamin B-6 analogue, increased the rate of dissociation of [3H]dexamethasone from HeLa S3 cytoplasmic glucocorticoid receptor complexes in vitro. This effect was achieved at millimolar concentrations of 5'-deoxypyridoxal, suggesting a low-affinity interaction of 5'-deoxypyridoxal with receptor. Loss of [3H]dexamethasone-receptor binding in the presence of 5'-deoxypyridoxal was pH dependent, and a plot of Kdiss vs. pH fit a simple sigmoidal titration curve with an inflection point at pH 7.8, suggesting that deprotonation of a single functional group on 5'-deoxypyridoxal increases Kdiss. Loss of [3H]dexamethasone binding in the presence or absence of unlabeled steroid also increased with pH, but no inflection point occurred over the range of pH tested. A titration of 5'-deoxypyridoxal indicated a pK of 7.94 for the pyridinium proton, suggesting deprotonation of the pyridinium nitrogen may account for the pH dependence of Kdiss of dexamethasone from receptor. 5'-Deoxypyridoxal also caused a decrease in nuclear [3H]-dexamethasone-receptor binding when incubated with whole HeLa S3 cells at 37 degrees C. Furthermore, 5'-deoxypyridoxal was effective in reducing nuclear binding of dexamethasone when added either simultaneously with [3H]dexamethasone or after achievement of equilibrium of steroid with receptor. The reduction in nuclear [3H]dexamethasone binding is highly specific for 5'-deoxypyridoxal. Several analogues of this compound, including 5'-deoxypyridoxamine, were ineffective. In addition, this effect was reversible following removal of extracellular 5'-deoxypyridoxal. Under these conditions, 5'-deoxypyridoxal was competitive with dexamethasone for binding to nuclear receptor, with KI = 8.1 X 10(-6) M. Scatchard plot analysis of dexamethasone-receptor binding in the presence or absence of 5'-deoxypyridoxal was consistent with an apparent reduced affinity of [3H]dexamethasone for receptor, which again suggests competitive interaction or allosteric interaction mediated dissociation. Glucocorticoids are known to stimulate alkaline phosphatase activity within HeLa S3 cells. In whole cell incubations, 5'-deoxypyridoxal was effective in reducing the dexamethasone-induced increase in alkaline phosphatase activity by 60% under conditions in which cell viability and cell growth were not affected.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Oct
pubmed:issn
0006-2960
pubmed:author
pubmed:issnType
Print
pubmed:day
26
pubmed:volume
21
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
5644-50
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed:year
1982
pubmed:articleTitle
5'-deoxypyridoxal interaction with dexamethasone receptor: a new probe for structure and function of steroid receptors.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.