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Transcriptional regulators bind to specific cis-regulatory sequences in the DNA to regulate gene transcription. These cis-regulatory sequences are very short, usually less than ten nucleotide pairs in length. The short length means that there is a high probability of the exact same sequence randomly occurring throughout the genome.  Since regulators can also bind to groups of similar sequences, this further increases the chances of random binding. Transcriptional regulators form dimers that...
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For successful DNA replication, the unwinding of double-stranded DNA must be accompanied by stabilization and protection of the separated single strands of the DNA. This crucial task is performed by single-strand DNA-binding (SSB) proteins. They bind to the DNA in a sequence-independent manner, which means that the nitrogenous bases of the DNA need not be present in a specific order for binding of SSB proteins to it. The binding of SSB proteins straightens single-stranded DNA (ssDNA) and makes...
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A Method for Selecting Structure-switching Aptamers Applied to a Colorimetric Gold Nanoparticle Assay
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Published on: February 28, 2015

Microarrays de receptores de estrógenos: subtipo selectivo de ligando de unión al ligando.

Sung Hoon Kim1, Anobel Tamrazi, Kathryn E Carlson

  • 1Department of Chemistry, University of Illinois, 600 South Matthews Avenue, Urbana, Illinois 61801, USA.

Journal of the American Chemical Society
|April 15, 2004
PubMed
Resumen

Desarrollamos un nuevo microarray receptor de hormonas nucleares para el estudio de los receptores de estrógeno (ERalpha-LBD y ERbeta-LBD). Este método de alto rendimiento evalúa de manera eficiente la unión de ligandos y puede adaptarse para otros receptores y el reclutamiento de correguladores.

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Área de la Ciencia:

  • La bioquímica es la bioquímica.
  • Biología Molecular Biología Molecular
  • Endocrinología Endocrinología.

Sus antecedentes:

  • Los receptores de hormonas nucleares juegan un papel crítico en los procesos celulares.
  • Los receptores de estrógenos (RE) son objetivos clave para comprender la regulación hormonal y la enfermedad.
  • Es esencial desarrollar métodos de alto rendimiento para estudiar las interacciones receptor-ligando.

Objetivo del estudio:

  • Para presentar el primer microarray receptor de hormonas nucleares.
  • Para ilustrar su aplicación utilizando los dominios de unión a los ligandos de los receptores de estrógenos alfa y beta (ERalpha-LBD y ERbeta-LBD).
  • Para demostrar su utilidad en el estudio de la unión de ligandos y la selectividad de subtipos.

Principales métodos:

  • Las proteínas (ERalpha-LBD y ERbeta-LBD) fueron impresas y unidas a las diapositivas de aldehído.
  • La eficiencia de la fijación se analizó en función del estado de unión a los ligandos (agonistas frente a antagonistas / no ligados).
  • La actividad de unión de ligandos se evaluó utilizando un conjugado estradiol-fluoroforo en ensayos de unión competitivos.

Principales resultados:

  • La eficiencia de fijación a las diapositivas fue significativamente mayor para los ER-LBDs no ligados o ligados a antagonistas en comparación con las formas ligadas a agonistas, lo que sugiere una unión específica de orientación.
  • Los ERs inmovilizados mantuvieron una significativa actividad de unión de ligandos.
  • Se determinó con éxito la unión específica y selectiva por subtipo de los ligandos a ERalpha-LBD y ERbeta-LBD.

Conclusiones:

  • Se ha establecido una nueva técnica de microarray de receptor de hormonas nucleares de alto rendimiento.
  • Este método permite una evaluación eficiente y selectiva de la unión del ligando a los receptores de estrógeno.
  • La plataforma es adaptable para el estudio de otros receptores de hormonas nucleares y el reclutamiento de correguladores.