Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Steroidogenic factor 1-DNA binding: a kinetic analysis using surface plasmon resonance.

D Bryan1, S J Aylwin, D J Newman

  • 1Department of Clinical Biochemistry, St Bartholomew's and the Royal London School of Medicine and Dentistry, Turner Street, London E1 2AD, UK.

Journal of Molecular Endocrinology
|May 27, 1999
PubMed
Summary

Steroidogenic factor-1 (SF-1) binds the glycoprotein hormone alpha-subunit gene

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Development and verification of an enzyme-linked immunosorbent assay for the quantification of toxoid A and toxoid B from Clostridioides difficile.

Journal of immunological methods·2020
Same author

BCG vaccination early in life does not improve COVID-19 outcome of elderly populations, based on nationally reported data.

Letters in applied microbiology·2020
Same author

Predicting pork loin intramuscular fat using computer vision system.

Meat science·2018
Same author

Technical developments of functional electrical stimulation to correct drop foot: sensing, actuation and control strategies.

Clinical biomechanics (Bristol, Avon)·2015
Same author

Depletion of penicillin G residues in heavy sows after intramuscular injection. Part I: tissue residue depletion.

Journal of agricultural and food chemistry·2014
Same author

Prediction of troponin-T degradation using color image texture features in 10d aged beef longissimus steaks.

Meat science·2013

Area of Science:

  • Molecular Endocrinology
  • Gene Regulation
  • Protein-DNA Interactions

Background:

  • Basal expression of the glycoprotein hormone alpha-subunit gene in pituitary gonadotrophs is regulated by a gonadotroph specific element (GSE).
  • The nuclear receptor steroidogenic factor-1 (SF-1) is known to bind the GSE, influencing gene expression.

Purpose of the Study:

  • To quantitatively analyze the binding kinetics of SF-1 to the GSE using surface plasmon resonance (SPR).
  • To investigate the impact of mutations within the GSE and flanking sequences on SF-1 binding affinity and kinetics.

Main Methods:

  • Surface plasmon resonance (SPR) was employed to determine association (kappa ass), dissociation (kappa diss), and affinity (KA) constants.
  • In vitro translated SF-1 protein and nuclear extracts from alphaT3-1 gonadotroph cells were used to assess binding to wild-type and mutant GSE oligonucleotides.

Related Experiment Videos

Main Results:

  • SF-1 exhibited a threefold higher affinity for the consensus GSE compared to a mutated GSE (GSEMUT), primarily due to increased association and decreased dissociation rates.
  • Binding specificity was confirmed by inhibition with anti-SF-1 antibody and excess unlabeled GSE.
  • Flanking sequences significantly reduced SF-1 affinity to both GSE and GSEMUT, mainly by decreasing the association rate.
  • Nuclear extracts showed slower dissociation rates than in vitro translated SF-1.

Conclusions:

  • SPR is a powerful tool for characterizing SF-1:GSE interactions and their modulation by sequence variations.
  • The study elucidates the kinetic parameters governing SF-1 binding to the glycoprotein hormone alpha-subunit gene promoter.
  • Findings highlight the importance of both the core binding site and flanking sequences in regulating SF-1-mediated gene expression.