NDLI: Dissecting the salt dependence of the Tus–Ter protein–DNA complexes by high-throughput differential scanning fluorimetry of a GFP-tagged Tus

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Toward mobile 3D visualization for structural biologists

Expanding the chemistry of fluorescent protein biosensors through genetic incorporation of unnatural amino acids

Network-based analysis of omics with multi-objective optimization

Cost-effective isobaric tagging for quantitative phosphoproteomics using DiART reagents

Gp10 based-thioetherification (10BASE_d-T) on a displaying library peptide of bacteriophage T7

Activity-based protein profiling of secreted cellulolytic enzyme activity dynamics in Trichoderma reesei QM6a, NG14, and RUT-C30

A fluorogenic probe for β-galactosidase activity imaging in living cells

The association of the cytoplasmic domains of interleukin 4 receptor alpha and interleukin 13 receptor alpha 2 regulates interleukin 4 signaling

Serum metabolic profiles reveal the effect of formoterol on cachexia in tumor-bearing mice

RNA-seq data analysis at the gene and CDS levels provides a comprehensive view of transcriptome responses induced by 4-hydroxynonenal

Binding structures and energies of the human neonatal Fc receptor with human Fc and its mutants by molecular modeling and dynamics simulations

Identification of biomarkers for unstable angina by plasma metabolomic profiling

Solvent interaction analysis of intrinsically disordered proteins in aqueous two-phase systems

Functional and pathway enrichment analysis for integrated regulatory network of high- and low-metastatic lung cancer

Artemisinin rewires the protein interaction network in cancer cells: network analysis, pathway identification, and target prediction

Transcriptomic study of ciprofloxacin resistance in Streptomyces coelicolor A3(2)

Effects of temperature on the dynamics of the LacI-TetR-CI repressilator

Design and biological evaluation of synthetic retinoids: probing length vs. stability vs. activity

Integrated proteomic and metabolomic analysis reveals the NADH-mediated TCA cycle and energy metabolism disorders based on a new model of chronic progressive heart failure

Dissecting the salt dependence of the Tus–Ter protein–DNA complexes by high-throughput differential scanning fluorimetry of a GFP-tagged Tus

Metabonomic investigations of age- and batch-related variations in female NMRI mice using proton nuclear magnetic resonance spectroscopy

PNA as a potential modulator of COL7A1 gene expression in dominant dystrophic epidermolysis bullosa: a physico-chemical study

Transfection activity and the mechanism of pDNA-complexes based on the hybrid of low-generation PAMAM and branched PEI-1.8k

Synergetic regulatory networks mediated by oncogene-driven microRNAs and transcription factors in serous ovarian cancer

Identification and comparative analysis of hepatitis C virus–host cell protein interactions

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Dissecting the salt dependence of the Tus–Ter protein–DNA complexes by high-throughput differential scanning fluorimetry of a GFP-tagged Tus

Content Provider	Royal Society of Chemistry (RSC)
Author	Schaeffer, Patrick M. Moreau, Morgane J. J.
Copyright Year	2013
Abstract	The analysis of the salt dependence of protein–DNA complexes provides useful information about the non-specific electrostatic and sequence-specific parameters driving complex formation and stability. The differential scanning fluorimetry of GFP-tagged protein (DSF-GTP) assay has been geared with an automatic Tm peak recognition system and was applied for the high-throughput (HT) determination of salt-induced effects on the GFP-tagged DNA replication protein Tus in complex with various Ter and Ter-lock sequences. The system was designed to generate two-dimensional heat map profiles of Tus-GFP protein stability allowing for a comparative study of the effect of eight increasing salt concentrations on ten different Ter DNA species at once. The data obtained with the new HT DSF-GTP allowed precise dissection of the non-specific electrostatic and sequence-specific parameters driving Tus–Ter and Tus–Ter-lock complex formation and stability. The major factor increasing the thermal resistance of Tus–Ter-lock complexes in high-salt is the formation of the TT-lock, e.g. a 10-fold higher Kspe was obtained for Tus-GFP:Ter-lockB than for Tus-GFP:TerB. It is anticipated that the system can be easily adapted for the study of other protein–DNA complexes.
Starting Page	3146
Ending Page	3154
Page Count	9
File Format	HTM / HTML PDF
ISSN	1742206X
Volume Number	9
Issue Number	12
Journal	Molecular BioSystems
DOI	10.1039/c3mb70426b
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Open
Subject Keyword	Fluorescence spectroscopy DNA replication DNA Kspe Protein Heat map
Content Type	Text
Resource Type	Article
Subject	Molecular Biology Biotechnology

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Differential Scanning Fluorimetry provides high throughput data on silk protein transitions.

Differential Tus-Ter binding and lock formation: implications for DNA replication termination in Escherichia coli.

Protocol for performing and optimizing differential scanning fluorimetry experiments.

Equilibrium, kinetic, and footprinting studies of the Tus-Ter protein-DNA interaction.

Mutations in the Escherichia coli Tus Protein Define a Domain Positioned Close to the DNA in the Tus-Ter Complex

Interplay between chromosomal architecture and termination of DNA replication in bacteria.

nanoDSF as screening tool for enzyme libraries and biotechnology development

Protein Microarray On-Demand: A Novel Protein Microarray System (2008)

High throughput challenges in molecular cell biology: the CELL MAP

Dissecting the salt dependence of the Tus–Ter protein–DNA complexes by high-throughput differential scanning fluorimetry of a GFP-tagged Tus

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