Characterization of DnaB–DnaG Interaction in M. tuberculosis Using Small-Angle X-ray Scattering-Based Dissociation Assay

A. Dayan; S. Ilic; B. Akabayov

doi:10.1002/cbic.202500289

Characterization of DnaB–DnaG Interaction in M. tuberculosis Using Small-Angle X-ray Scattering-Based Dissociation Assay

A. Dayan, S. Ilic, B. Akabayov

Research output: Contribution to journal › Article › peer-review

Abstract

The complex interactions between helicase and primase, two key components of the replisome involved in DNA replication in Mycobacterium tuberculosis are studied. Utilizing purified, complementary domains of these proteins, a surface plasmon resonance (SPR) analysis and a cross-linking assay to characterize their binding dynamics are employed. The SPR analysis reveals a binding dissociation constant of 0.21 ± 0.08 μM, and the cross-linking assay suggests the possible formation of a heterodimer species. Importantly, a small-angle X-ray scattering dissociation assay to study the dynamic interactions between the proteins in solution is utilized. The findings provide new opportunities for targeted therapeutic strategies aimed at DNA replication in M. tuberculosis by revealing the structural interplay between helicase and primase.

Original language	English
Article number	e202500289
Journal	ChemBioChem
Volume	26
Issue number	14
DOIs	https://doi.org/10.1002/cbic.202500289
State	Published - 18 Jul 2025

Keywords

DnaB
DnaG
protein–protein interactions
small-angle X-ray scattering dissociation assay

ASJC Scopus subject areas

Biochemistry
Molecular Medicine
Molecular Biology
Organic Chemistry

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1002/cbic.202500289

Cite this

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title = "Characterization of DnaB–DnaG Interaction in M. tuberculosis Using Small-Angle X-ray Scattering-Based Dissociation Assay",

abstract = "The complex interactions between helicase and primase, two key components of the replisome involved in DNA replication in Mycobacterium tuberculosis are studied. Utilizing purified, complementary domains of these proteins, a surface plasmon resonance (SPR) analysis and a cross-linking assay to characterize their binding dynamics are employed. The SPR analysis reveals a binding dissociation constant of 0.21 ± 0.08 μM, and the cross-linking assay suggests the possible formation of a heterodimer species. Importantly, a small-angle X-ray scattering dissociation assay to study the dynamic interactions between the proteins in solution is utilized. The findings provide new opportunities for targeted therapeutic strategies aimed at DNA replication in M. tuberculosis by revealing the structural interplay between helicase and primase.",

keywords = "DnaB, DnaG, protein–protein interactions, small-angle X-ray scattering dissociation assay",

author = "A. Dayan and S. Ilic and B. Akabayov",

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doi = "10.1002/cbic.202500289",

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AU - Dayan, A.

AU - Ilic, S.

AU - Akabayov, B.

PY - 2025/7/18

Y1 - 2025/7/18

N2 - The complex interactions between helicase and primase, two key components of the replisome involved in DNA replication in Mycobacterium tuberculosis are studied. Utilizing purified, complementary domains of these proteins, a surface plasmon resonance (SPR) analysis and a cross-linking assay to characterize their binding dynamics are employed. The SPR analysis reveals a binding dissociation constant of 0.21 ± 0.08 μM, and the cross-linking assay suggests the possible formation of a heterodimer species. Importantly, a small-angle X-ray scattering dissociation assay to study the dynamic interactions between the proteins in solution is utilized. The findings provide new opportunities for targeted therapeutic strategies aimed at DNA replication in M. tuberculosis by revealing the structural interplay between helicase and primase.

AB - The complex interactions between helicase and primase, two key components of the replisome involved in DNA replication in Mycobacterium tuberculosis are studied. Utilizing purified, complementary domains of these proteins, a surface plasmon resonance (SPR) analysis and a cross-linking assay to characterize their binding dynamics are employed. The SPR analysis reveals a binding dissociation constant of 0.21 ± 0.08 μM, and the cross-linking assay suggests the possible formation of a heterodimer species. Importantly, a small-angle X-ray scattering dissociation assay to study the dynamic interactions between the proteins in solution is utilized. The findings provide new opportunities for targeted therapeutic strategies aimed at DNA replication in M. tuberculosis by revealing the structural interplay between helicase and primase.

KW - DnaB

KW - DnaG

KW - protein–protein interactions

KW - small-angle X-ray scattering dissociation assay

UR - https://www.scopus.com/pages/publications/105009491691

U2 - 10.1002/cbic.202500289

DO - 10.1002/cbic.202500289

M3 - Article

C2 - 40497748

AN - SCOPUS:105009491691

SN - 1439-4227

VL - 26

JO - ChemBioChem

JF - ChemBioChem

IS - 14

M1 - e202500289

ER -

Characterization of DnaB–DnaG Interaction in M. tuberculosis Using Small-Angle X-ray Scattering-Based Dissociation Assay

Abstract

Keywords

ASJC Scopus subject areas

UN SDGs

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