FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W2076328350> ?p ?o ?g. }

• W2076328350 endingPage "178" @default.
• W2076328350 startingPage "169" @default.
• W2076328350 abstract "The expression of high levels of full-length human estrogen receptor alpha (hERalpha) in Escherichia coli has proven difficult. We found that expression of the ER DNA binding domain is highly toxic to E. coli, resulting in rapid loss of the expression plasmid. Using a tightly regulated arabinose expression system and the antibiotic Timentin, we were able to overcome ER toxicity and express substantial levels of ER. The expressed ER exhibited protease cleavage at a single site near the N-terminus of the hinge region. Of the many measures we tested to eliminate ER cleavage, only addition of carbonyl cyanide m-chlorophenyl-hydrazone (CCCP), an uncoupler of oxidative phosphorylation, completely blocked intracellular proteolysis of the ER. Using CCCP and our expression methods, full-length FLAG epitope-tagged hERalpha (fER) was expressed in E. coli at approximately 1 mg/l. The fER was purified to homogeneity in a single step by immunoaffinity chromatography with anti-FLAG monoclonal antibody. Purified full-length bacterial fER binds 17beta-estradiol with the same affinity as hER expressed in human cells (K(D) approximately 0.5 nM). At high concentrations of fER (20 nM), a bell-shaped estrogen binding curve with a Hill coefficient of 1.7 was seen. Bacterially-expressed fER exhibits a reduced affinity for the estrogen response element (ERE). Anti-FLAG antibody restores high affinity binding of the fER to the ERE, suggesting that impaired dimerization may be responsible for the reduced affinity of bacterially-expressed fER for the ERE. The use of Timentin and CCCP may provide a general method for high level bacterial expression of steroid/nuclear receptors and other proteins important in hormone action." @default.
• W2076328350 created "2016-06-24" @default.
• W2076328350 creator A5030190949 @default.
• W2076328350 creator A5061599949 @default.
• W2076328350 creator A5066830496 @default.
• W2076328350 creator A5067562997 @default.
• W2076328350 date "2000-11-01" @default.
• W2076328350 modified "2023-10-18" @default.
• W2076328350 title "High level expression of full-length estrogen receptor in Escherichia coli is facilitated by the uncoupler of oxidative phosphorylation, CCCP" @default.
• W2076328350 cites W1489022720 @default.
• W2076328350 cites W1549507201 @default.
• W2076328350 cites W1857981040 @default.
• W2076328350 cites W1927136478 @default.
• W2076328350 cites W1928071293 @default.
• W2076328350 cites W1966568300 @default.
• W2076328350 cites W1967286436 @default.
• W2076328350 cites W1979439941 @default.
• W2076328350 cites W1985006541 @default.
• W2076328350 cites W1995534157 @default.
• W2076328350 cites W1999999697 @default.
• W2076328350 cites W2001269969 @default.
• W2076328350 cites W2003580491 @default.
• W2076328350 cites W2004356664 @default.
• W2076328350 cites W2007203662 @default.
• W2076328350 cites W2008398027 @default.
• W2076328350 cites W2014107736 @default.
• W2076328350 cites W2027654568 @default.
• W2076328350 cites W2032250517 @default.
• W2076328350 cites W2032552404 @default.
• W2076328350 cites W2041852535 @default.
• W2076328350 cites W2049416739 @default.
• W2076328350 cites W2056188223 @default.
• W2076328350 cites W2059709861 @default.
• W2076328350 cites W2072832822 @default.
• W2076328350 cites W2094230077 @default.
• W2076328350 cites W2108593941 @default.
• W2076328350 cites W2114332227 @default.
• W2076328350 cites W2128787958 @default.
• W2076328350 cites W2132185105 @default.
• W2076328350 cites W2133293679 @default.
• W2076328350 cites W2142365365 @default.
• W2076328350 cites W2155755249 @default.
• W2076328350 cites W2160084442 @default.
• W2076328350 cites W2199565660 @default.
• W2076328350 cites W2314479559 @default.
• W2076328350 cites W2615633554 @default.
• W2076328350 cites W4233578116 @default.
• W2076328350 cites W4237911804 @default.
• W2076328350 cites W4239735288 @default.
• W2076328350 cites W4241546774 @default.
• W2076328350 cites W4245192752 @default.
• W2076328350 cites W4254810143 @default.
• W2076328350 doi "https://doi.org/10.1016/s0960-0760(00)00120-5" @default.
• W2076328350 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/11162922" @default.
• W2076328350 hasPublicationYear "2000" @default.
• W2076328350 type Work @default.
• W2076328350 sameAs 2076328350 @default.
• W2076328350 citedByCount "9" @default.
• W2076328350 countsByYear W20763283502012 @default.
• W2076328350 countsByYear W20763283502013 @default.
• W2076328350 countsByYear W20763283502022 @default.
• W2076328350 crossrefType "journal-article" @default.
• W2076328350 hasAuthorship W2076328350A5030190949 @default.
• W2076328350 hasAuthorship W2076328350A5061599949 @default.
• W2076328350 hasAuthorship W2076328350A5066830496 @default.
• W2076328350 hasAuthorship W2076328350A5067562997 @default.
• W2076328350 hasConcept C104317684 @default.
• W2076328350 hasConcept C107824862 @default.
• W2076328350 hasConcept C11960822 @default.
• W2076328350 hasConcept C121608353 @default.
• W2076328350 hasConcept C153911025 @default.
• W2076328350 hasConcept C179247698 @default.
• W2076328350 hasConcept C181199279 @default.
• W2076328350 hasConcept C185592680 @default.
• W2076328350 hasConcept C51445715 @default.
• W2076328350 hasConcept C530470458 @default.
• W2076328350 hasConcept C54355233 @default.
• W2076328350 hasConcept C547475151 @default.
• W2076328350 hasConcept C55493867 @default.
• W2076328350 hasConcept C84606932 @default.
• W2076328350 hasConcept C86803240 @default.
• W2076328350 hasConceptScore W2076328350C104317684 @default.
• W2076328350 hasConceptScore W2076328350C107824862 @default.
• W2076328350 hasConceptScore W2076328350C11960822 @default.
• W2076328350 hasConceptScore W2076328350C121608353 @default.
• W2076328350 hasConceptScore W2076328350C153911025 @default.
• W2076328350 hasConceptScore W2076328350C179247698 @default.
• W2076328350 hasConceptScore W2076328350C181199279 @default.
• W2076328350 hasConceptScore W2076328350C185592680 @default.
• W2076328350 hasConceptScore W2076328350C51445715 @default.
• W2076328350 hasConceptScore W2076328350C530470458 @default.
• W2076328350 hasConceptScore W2076328350C54355233 @default.
• W2076328350 hasConceptScore W2076328350C547475151 @default.
• W2076328350 hasConceptScore W2076328350C55493867 @default.
• W2076328350 hasConceptScore W2076328350C84606932 @default.
• W2076328350 hasConceptScore W2076328350C86803240 @default.
• W2076328350 hasIssue "4" @default.
• W2076328350 hasLocation W20763283501 @default.

• next