Local conformations affect the histidine tag-Ni&lt;sup&gt;2+&lt;/sup&gt; binding affinity of BinA and BinB proteins

Tharad, Sudarat; Tangsongcharoen, Chontida; Boonserm, Panadda; Toca-Herrera, José L.; Srisucharitpanit, Kanokporn

doi:10.3934/biophy.2020011

Cited by 2 publications

(3 citation statements)

References 19 publications

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“…Because 6H-GFP is much larger, its translational and rotational diffusion is reduced which facilitates the formation of the second bond. Different molecular interactions between the bilayer and His-tagged molecules might also play a role in the reduced affinity of 6H-FITC compared to 6H-GFP as proposed for other types of His-tagged proteins 49 .…”

Section: Binding Of 6h-fitc To Guv Membranesmentioning

confidence: 92%

Binding of His-tagged fluorophores to lipid bilayers of giant vesicles†

Pramanik

Steinkühler²,

Dimova

et al. 2022

Preprint

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His-tagged molecules can be attached to lipid bilayers via certain anchor lipids, a method that has been widely used for the biofunctionalization of membranes and vesicles. To measure the coverage by the membrane-bound molecules, it is useful to study molecules that are fluorescent as well. Here, we use two such molecules, green fluorescence protein (GFP) and green-fluorescent fluorescin isothiocyanate (FITC), both of which are tagged with a chain of six histidines that bind to achor lipids within the bilayers. This His-tag is much smaller in size than the GFP molecule but somewhat larger than the FITC dye. The lipid bilayers form giant unilamellar vesicles (GUVs), the behavior of which can be directly observed in the optical microscope. Several protocols for the preparation of GUVs have been developed. We apply and compare three well-established protocols based on polyvinyl alcohol (PVA) hydrogel swelling, electroformation on platinum wires, and electroformation on indium tin oxide (ITO) glass. For the same nanomolar concentration in the exterior solution, the coverage by His-tagged FITC is much lower than the one by His-tagged GFP. However, for both GFP and FITC, we find that the binding of the His-tagged molecules to the anchor lipids depends strongly on the preparation method. The highest binding affinitiy is obtained for electroformation on platinum wires. PVA gel swelling gives rise to a somewhat smaller binding affinity whereas electroformation on ITO glass leads to essentially no binding. Furthermore, the binding affinitiy is also observed to depend on the pH of the aqueous solution, with a relatively weak and strong pH-dependence for His-tagged GFP and His-tagged FITC, respectively.

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Section: Binding Of 6h-fitc To Guv Membranesmentioning

confidence: 92%

Binding of His-tagged fluorophores to lipid bilayers of giant vesicles†

Pramanik

Steinkühler²,

Dimova

et al. 2022

Preprint

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“…QCM-D experiments were performed to study the lipid bilayer formation and its interaction with the binary toxin, imitating the binding of binary toxin to the susceptible cell membrane. 81 It is particularly useful for identifying different types of biomolecules based on the target-dependent adhesion fingerprints of S. cerevisiae and E. coli. 12,82 Not only does it present a native-like environment, the method also facilitates the discovery of new receptors of emerging strains of highly mutating viruses and the development of new antiviral drugs.…”

Section: Characterizations Using Qcm-dmentioning

confidence: 99%

“…Camesano and co-workers , used QCM-D to determine the concentration- and time-dependent changes in mass adsorption, film rigidity, thickness, and density of zwitterionic bilayers upon interaction with an antimicrobial peptide LL37 at concentrations representing its antimicrobial (low concentrations) and cytotoxic activities (high concentrations). QCM-D experiments were performed to study the lipid bilayer formation and its interaction with the binary toxin, imitating the binding of binary toxin to the susceptible cell membrane . It is particularly useful for identifying different types of biomolecules based on the target-dependent adhesion fingerprints of S. cerevisiae and E. coli . , Not only does it present a native-like environment, the method also facilitates the discovery of new receptors of emerging strains of highly mutating viruses and the development of new antiviral drugs. , To date, the investigation of cell–molecule interactions has been expanded to cover a large variety of molecules, including carbohydrates, cytoskeleton-targeting agents, proteins, RGD ligands, cytomorphic agents, etc.…”

Section: An Overview Of Biomacromolecular Characterizations Using Qcm-dmentioning

confidence: 99%

Biomacromolecular Characterizations Using State-of-the-Art Quartz Crystal Microbalance with Dissipation

Wei,

Shao,

Qiao

et al. 2023

Anal. Chem.

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Biomolecular characterization is essential in fields such as drug discovery, glycomics, and cell biology. This feature article focuses on the experimental use of quartz crystal microbalance with dissipation (QCM-D) as a powerful analytical technique to probe biological events ranging from biomacromolecular interactions and conformational changes of biomacromolecules to surface immobilization of biomacromolecules and cell morphological changes.

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Local conformations affect the histidine tag-Ni<sup>2+</sup> binding affinity of BinA and BinB proteins

Cited by 2 publications

References 19 publications

Binding of His-tagged fluorophores to lipid bilayers of giant vesicles†

Binding of His-tagged fluorophores to lipid bilayers of giant vesicles†

Biomacromolecular Characterizations Using State-of-the-Art Quartz Crystal Microbalance with Dissipation

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