Evaluating Electrostatic Contributions to Binding with the Use of Protein Charge Ladders

@article{Gao1996EvaluatingEC,
  title={Evaluating Electrostatic Contributions to Binding with the Use of Protein Charge Ladders},
  author={J Gao and Mathai Mammen and George M. Whitesides},
  journal={Science},
  year={1996},
  volume={272},
  pages={535 - 537}
}
Electrostatic interactions between charges on ligands and charges on proteins that are remote from the binding interface can influence the free energy of binding (ΔGb). The binding affinities between charged ligands and the members of a charge ladder of bovine carbonic anhydrase (CAII) constructed by random acetylation of the amino groups on its surface were measured by affinity capillary electrophoresis (ACE). The values of ΔGb derived from this analysis correlated approximately linearly with… 
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66 Citations
Background Citations
8
Methods Citations
5
Results Citations
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66 Citations

Citation Type

Citation Type

Using charge ladders and capillary electrophoresis to measure the charge, size, and electrostatic interactions of proteins.
TLDR
This chapter provides an overview of protein charge ladders--collections of protein derivatives that differ in charge--and capillary electrophoresis (CE), a useful biophysical tool for measuring the net charge of proteins and the role of electrostatics in biochemical processes involving proteins.
Why are proteins charged? Networks of charge-charge interactions in proteins measured by charge ladders and capillary electrophoresis.
TLDR
By studying the influence of charge on the properties of proteins using charge ladders, it is possible to estimate the net charge and hydrodynamic radius and to infer the role of charged residues in ligand binding and protein folding.
Electrostatics and packing in biomolecules : accounting for conformational change in protein folding and binding
TLDR
A measure of electrostatic complementarity was developed and applied to myoglobin—both to known sequences and to hypothetical chimeric myoglobin sequences, and performed better than other readily available measures of complementarity when myoglobin homology models were evaluated.
Protein charge ladders: a new technique for studying electrostatic interactions in ultrafiltration systems
Competitive adsorption of model charged proteins: the effect of total charge and charge distribution.
Direct molecular level measurements of the electrostatic properties of a protein surface.
TLDR
Although this study focuses on the pH-dependence of surface electrostatics, this direct approach to probing the electrostatic features of proteins is applicable to investigations of any perturbations that alter the charge distribution of the surfaces of immobilized molecules.
Affinity capillary electrophoresis: A physical‐organic tool for studying interactions in biomolecular recognition
TLDR
This review emphasizes ACE experiments performed with two model systems: bovine carbonic anhydrase II (BCA II) with arylsulfonamide ligands and vancomycin (Van), a glycopeptide antibiotic, with D‐Ala‐D‐ Ala (DADA)‐based peptidyl ligands, and variations of ACE experiments have been used to estimate the charge of Van and of proteins in solution.
Increasing the net charge and decreasing the hydrophobicity of bovine carbonic anhydrase decreases the rate of denaturation with sodium dodecyl sulfate.
TLDR
The kinetics of denaturation in solutions containing SDS of the protein charge ladders generated with acetic and hexanoic anhydrides are reported, with the proteins with an intermediate number of modifications being the most stable to denaturation by SDS.
Modulation of protein stability and aggregation properties by surface charge engineering.
TLDR
When the protein properties such as fluorescent activity, folding rate and kinetic stability were assessed, it was found the possibility that the protein stability can be modulated independently of activity and folding by engineering protein surface charges.
Neutralizing positive charges at the surface of a protein lowers its rate of amide hydrogen exchange without altering its structure or increasing its thermostability.
TLDR
This paper combines two techniques--mass spectrometry and protein charge ladders--to examine the relationship between the surface charge and hydrophobicity of a representative globular protein and its rate of amide hydrogen-deuterium (H/D) exchange and suggests that the electrostatic charge is more important than the hydrophobia of surface groups in determining the rate of H/D exchange.
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  • 1996
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