Nucleotide Binding

Nucleotide binding is the reversible recognition of nucleotides by proteins, nucleic acids, or other biomolecules, enabling cells to use these molecules as energy sources, genetic building blocks, and regulatory signals. Binding occurs when a nucleotide’s base, sugar, and phosphate groups form complementary hydrogen bonds, electrostatic interactions, and hydrophobic contacts with a binding site; magnesium ions may also coordinate phosphate groups and stabilize the complex. In biochemistry, nucleotide binding controls the activity of enzymes, transporters, and molecular switches such as ATP- and GTP-binding proteins. Studying these interactions helps explain energy transfer, signal transduction, DNA and RNA metabolism, and the action of nucleotide-targeting drugs.

Nucleotide Binding - Related Videos

Research

JoVE EoE - Assay Techniques

Differential Radial Capillary Action of Ligand Assay: A High-Throughput Technique to Identify Bacterial Nucleotide Second Messenger-Binding Intracellular Proteins

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2025

This video demonstrates the differential radial capillary action of ligand assay — a technique for systematic high-throughput screening of intracellular nucleotide second messenger-binding proteins. This technique allows for direct binding of the messengers to the overexpressed target protein in the cell lysate — bypassing the need for protein purification.

Research

JoVE Journal - Biochemistry
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Measuring Nucleotide Binding to Intact, Functional Membrane Proteins in Real Time

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Cited by 4 •

2021

This protocol presents a method for measuring adenine nucleotide binding to receptors in real time in a cellular environment. Binding is measured as Förster resonance energy transfer (FRET) between trinitrophenyl nucleotide derivatives and protein labeled with a non-canonical, fluorescent amino acid.

Competition Binding Assay to Study Competing GTPase-Binding Protein Partners

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2025

This video demonstrates a competition assay to study GTPase-binding protein partners. Utilizing nucleotide-bound GTPase protein immobilized on magnetic beads, the competitive binding between two interacting protein partners for the same binding site on the GTPase can be studied to assess the binding affinities of the protein partners.

Education

JoVE Core - Molecular Biology

Cooperative Binding of Transcription Regulators

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2020

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...

Research

JoVE Journal - Medicine

Genotyping Single Nucleotide Polymorphisms in the Mitochondrial Genome by Pyrosequencing

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Cited by 5 •

2023

Pyrosequencing assays enable the robust and rapid genotyping of mitochondrial DNA single nucleotide polymorphisms in heteroplasmic cells or tissues.

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