Binding Kinetics I Conformational Changes
|Affinity, dose response||KD, IC50, fM sensitivity||Protein diameter||DH, ΔDH = 0.1 nm|
|Kinetics||kON, kOFF||Thermodynamics||ΔH, ΔS, ΔG|
|Avidity, bispecifics||Two-color detection||Nucleic acid enzyme activity||kCAT, KM, kEXO|
|Conformational changes||% friction change|
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Browse through the latest publications.
Paper in Environmental Toxicology and Pharmacology I 03 September 2020
In this publication, Delannoy at al. take advantage of switchSENSE® technology for studying protein – small molecules interaction, measuring the binding affinity of chlordecone and its metabolite chlordecol (CLD and CLD-OH) to isolated human lipoproteins (LDL and HDL).
CLD is known to be one of the most frequently detected chlorinated pesticide, which has a huge impact on health, including increased incidence of prostate cancer as well as neurodevelopment disorders. CLD accumulates in liver, binds to lipoproteins (e.g. HDL, LDL) and circulates in the blood.
This work provides useful information about the specificity of interaction of CLD with LDL and CLD-OH with HDL, obtained using switchSENSE®, allowing the development of a distribution model of CLD and CLD-OH in human tissues and a better understanding of CLD bioavailabity in humans.
Paper in Analytical Biochemistry I 19 July 2020
Nucleotide binding kinetics and conformational change analysis of tissue transglutaminase with switchSENSE
In this publication, we introduce switchSENSE® as a versatile tool for TG2 characterization and provide novel insights into protein conformation as well as analyte binding kinetics. For the first time, we succeeded in measuring the kinetic rate constants and affinities (kon, koff, KD) for guanosine nucleotides (GMP, GDP, GTP, GTPγS). Further, the conformational changes induced by GDP, Ca2+ and the covalent inhibitor Z-DON were observed by changes in TG2’s hydrodynamic diameter. We confirmed the well-known compaction by guanosine nucleotides and extension by Ca2+, and provide evidence for TG2 conformations so far not described by structural analysis. Moreover, we analyze the influence of the peptidic Z-DON inhibitor and the R580A mutation on the conformational responsiveness of TG2 to its natural effectors. In summary, this work shows how the combination of structural and kinetic information obtained by switchSENSE® opens new perspectives for the characterization of conformationally active proteins and their interactions with ligands, e.g. potential drug candidates.
Paper in Scientific Reports I 09 June 2020
The trimer to monomer transition of Tumor Necrosis Factor-Alpha is a dynamic process that is significantly altered by therapeutic antibodies
In this paper, we demonstrate the power of switchSENSE® for size analysis and antibody binding kinetics. In certain inflammatory diseases, such as rheumatoid arthritis, TNF-α is upregulated and forms bioactive homotrimers that promote inflammation. This bioactivation that was previously reported to occur over hours is seen to occur in only minutes when measured in real-time using switchSENSE’s dynamic measurement mode. The kinetics of TNF-α targeting therapeutic antibodies, including HUMIRA, are also observed, including the long dissociation (measured over 30,000 sec!) of TNF-α from these tight binders. This novel approach also sheds some light on the strong differences between these antibodies that were previously thought to have the same mode of action.