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

A robust method for determining DNA binding constants using capillary zone electrophoresis

C Li1, L M Martin

  • 1College of Pharmacy, University of Rhode Island, 41 Lower College Road, Kingston, Rhode Island 02881-0809, USA.

Analytical Biochemistry
|September 29, 1998
PubMed
Summary
This summary is machine-generated.

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

Neuropeptide Y and tuberoinfundibular dopamine activities are altered during lactation: role of prolactin.

Endocrinology·1999
Same author

Blood pressure and heart rate in the ovine fetus: ontogenic changes and effects of fetal adrenalectomy.

The American journal of physiology·1999
Same author

Characteristics of delayed excretion of flavonoids in human urine after administration of Shosaiko-to, a herbal medicine.

Biological & pharmaceutical bulletin·1999
Same author

A long terminal repeat of the human endogenous retrovirus ERV-9 is located in the 5' boundary area of the human beta-globin locus control region.

Genomics·1999
Same author

Evaluation of a mucoadhesive buccal patch for delivery of peptides: in vitro screening of bioadhesion.

Drug development and industrial pharmacy·1999
Same author

A targeted disruption of the murine Brca1 gene causes gamma-irradiation hypersensitivity and genetic instability.

Oncogene·1999
Same journal

Lysozyme assay using a rationally designed GN4G2 substrate with coupled β-glucosidase reaction.

Analytical biochemistry·2026
Same journal

The long run: A tribute to Arthur Joseph Lawrence Cooper.

Analytical biochemistry·2026
Same journal

Evaluation of a method for affinity measurement using solution equilibrium titration with magnetic beads.

Analytical biochemistry·2026
Same journal

Metabolomics approach using UHPLC/QE-MS for the mechanism of He Xue Ming Mu tablets on non-proliferative diabetic retinopathy.

Analytical biochemistry·2026
Same journal

UniRES-GO: Unified residue-level early fusion of sequence and predicted structure for protein function prediction.

Analytical biochemistry·2026
Same journal

IgG detection by enzyme-linked mass spectrometric assay versus color, fluorescent, ECL in buffer and serum.

Analytical biochemistry·2026
See all related articles

This study used capillary electrophoresis to measure DNA-binding affinities of novel tetrapeptides. The designed peptides showed a wide range of DNA-binding constants, from 10^2 to 10^6 M^-1.

Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Molecular Biology

Background:

  • Calf thymus DNA is a common model for studying DNA-protein interactions.
  • Tetrapeptides with unnatural amino acids offer unique structural and functional properties.
  • Capillary Zone Electrophoresis (CZE) is a powerful separation technique for analyzing molecular interactions.

Purpose of the Study:

  • To quantify the binding constants between calf thymus DNA and a library of designed tetrapeptides.
  • To investigate the DNA-binding affinities of peptides synthesized with unnatural amino acids containing thiazole rings.
  • To establish a method for determining DNA-peptide binding using CZE.

Main Methods:

  • Capillary Zone Electrophoresis (CZE) with on-line UV detection was employed.

Related Experiment Videos

  • Equilibrium mixtures of DNA and tetrapeptides were prepared at 8°C for 12 hours.
  • Scatchard plot analysis of unbound tetrapeptide concentrations determined binding constants.

    • Main Results:

    • CZE successfully separated free tetrapeptides from DNA-peptide complexes.
    • UV absorbance of unbound tetrapeptide decreased with increasing DNA concentration.
    • DNA-binding affinities ranged from 10^2 to 10^6 M^-1 for the tested tetrapeptides.

    Conclusions:

    • The developed CZE method effectively measures DNA-peptide binding constants.
    • The designed tetrapeptides exhibit a broad spectrum of DNA-binding affinities.
    • Unnatural amino acid incorporation influences peptide-DNA interaction strength.