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The Role of Electrostatic Interactions in Binding of Histone H3K4me2/3 to the Sgf29 Tandem Tudor Domain.

Pieters BJ, Meulenbroeks E, Belle R, Mecinović J - PLoS ONE (2015)

Bottom Line: Several reader domain proteins that specifically recognize methyllysine-containing histones contain the negatively-charged aspartate or glutamate residues as part of the aromatic cage.Small uncharged and large aromatic substitutions on the Asp266 site resulted in a significant decrease in binding affinities for both H3K4me3 and H3K4me2, demonstrating the role of the negative charge of Asp266 in the readout process by Sgf29.This study emphasizes the essential contribution of electrostatic interactions to the overall binding affinity, and reveals that the underlying mechanisms for the recognition of Kme2/3 depend on the composition and arrangement of the aromatic cage.

View Article: PubMed Central - PubMed

Affiliation: Institute of Molecules and Materials, Radboud University Nijmegen, Nijmegen, The Netherlands.

ABSTRACT
Several reader domain proteins that specifically recognize methyllysine-containing histones contain the negatively-charged aspartate or glutamate residues as part of the aromatic cage. Herein, we report thermodynamic analyses for the recognition of histone H3K4me3 and H3K4me2 by the tandem tudor domain of Sgf29 and its recognition site variants. Small uncharged and large aromatic substitutions on the Asp266 site resulted in a significant decrease in binding affinities for both H3K4me3 and H3K4me2, demonstrating the role of the negative charge of Asp266 in the readout process by Sgf29. This study emphasizes the essential contribution of electrostatic interactions to the overall binding affinity, and reveals that the underlying mechanisms for the recognition of Kme2/3 depend on the composition and arrangement of the aromatic cage.

No MeSH data available.


ITC experiments showing binding of A) ARTKme3QTAGKS and B) ARTKme3QTA to WT Sgf29.Thermodynamics of binding for A) Kd = 4.0 ± 0.6 μM, ΔG° = - 7.4 ± 0.1 kcal mol-1, ΔH° = - 8.0 ± 0.1 kcal mol-1,-TΔS° = 0.6 ± 0.1 kcal mol-1 and for B) Kd = 9.0 ± 0.5 μM, ΔG° = - 6.9 ± 0.1 kcal mol-1, ΔH° = - 7.8 ± 0.1 kcal mol-1,-TΔS° = 0.9 ± 0.1 kcal mol-1.
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pone.0139205.g006: ITC experiments showing binding of A) ARTKme3QTAGKS and B) ARTKme3QTA to WT Sgf29.Thermodynamics of binding for A) Kd = 4.0 ± 0.6 μM, ΔG° = - 7.4 ± 0.1 kcal mol-1, ΔH° = - 8.0 ± 0.1 kcal mol-1,-TΔS° = 0.6 ± 0.1 kcal mol-1 and for B) Kd = 9.0 ± 0.5 μM, ΔG° = - 6.9 ± 0.1 kcal mol-1, ΔH° = - 7.8 ± 0.1 kcal mol-1,-TΔS° = 0.9 ± 0.1 kcal mol-1.

Mentions: The involvement of the potential salt-bridge between the negatively-charged D266 and positively-charged R8 of 10-mer H3K4me3 and H3K4me2 peptides was also examined (PDB ID: 3ME9, 3MET). 10-mer H3K4me3 peptide that contains G instead of R at position 8 bound to WT Sgf29 with a Kd of 4 μM, similar to the native 10-mer peptide (Fig 6A). In addition, 7-mer peptide ARTKme3QTA that lacks the last three residues, including the eighth residue R8, bound to WT Sgf29 with Kd value of 9 μM, demonstrating the 3-fold decrease of the binding affinity relative to 10-mer H3K4me3 peptide (Fig 6B). These data suggest that it is unlikely that potential D266-R8 salt-bridge contributes to a significant extent to the overall favorable association between H3K4m2/3 and Sgf29.


The Role of Electrostatic Interactions in Binding of Histone H3K4me2/3 to the Sgf29 Tandem Tudor Domain.

Pieters BJ, Meulenbroeks E, Belle R, Mecinović J - PLoS ONE (2015)

ITC experiments showing binding of A) ARTKme3QTAGKS and B) ARTKme3QTA to WT Sgf29.Thermodynamics of binding for A) Kd = 4.0 ± 0.6 μM, ΔG° = - 7.4 ± 0.1 kcal mol-1, ΔH° = - 8.0 ± 0.1 kcal mol-1,-TΔS° = 0.6 ± 0.1 kcal mol-1 and for B) Kd = 9.0 ± 0.5 μM, ΔG° = - 6.9 ± 0.1 kcal mol-1, ΔH° = - 7.8 ± 0.1 kcal mol-1,-TΔS° = 0.9 ± 0.1 kcal mol-1.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4589357&req=5

pone.0139205.g006: ITC experiments showing binding of A) ARTKme3QTAGKS and B) ARTKme3QTA to WT Sgf29.Thermodynamics of binding for A) Kd = 4.0 ± 0.6 μM, ΔG° = - 7.4 ± 0.1 kcal mol-1, ΔH° = - 8.0 ± 0.1 kcal mol-1,-TΔS° = 0.6 ± 0.1 kcal mol-1 and for B) Kd = 9.0 ± 0.5 μM, ΔG° = - 6.9 ± 0.1 kcal mol-1, ΔH° = - 7.8 ± 0.1 kcal mol-1,-TΔS° = 0.9 ± 0.1 kcal mol-1.
Mentions: The involvement of the potential salt-bridge between the negatively-charged D266 and positively-charged R8 of 10-mer H3K4me3 and H3K4me2 peptides was also examined (PDB ID: 3ME9, 3MET). 10-mer H3K4me3 peptide that contains G instead of R at position 8 bound to WT Sgf29 with a Kd of 4 μM, similar to the native 10-mer peptide (Fig 6A). In addition, 7-mer peptide ARTKme3QTA that lacks the last three residues, including the eighth residue R8, bound to WT Sgf29 with Kd value of 9 μM, demonstrating the 3-fold decrease of the binding affinity relative to 10-mer H3K4me3 peptide (Fig 6B). These data suggest that it is unlikely that potential D266-R8 salt-bridge contributes to a significant extent to the overall favorable association between H3K4m2/3 and Sgf29.

Bottom Line: Several reader domain proteins that specifically recognize methyllysine-containing histones contain the negatively-charged aspartate or glutamate residues as part of the aromatic cage.Small uncharged and large aromatic substitutions on the Asp266 site resulted in a significant decrease in binding affinities for both H3K4me3 and H3K4me2, demonstrating the role of the negative charge of Asp266 in the readout process by Sgf29.This study emphasizes the essential contribution of electrostatic interactions to the overall binding affinity, and reveals that the underlying mechanisms for the recognition of Kme2/3 depend on the composition and arrangement of the aromatic cage.

View Article: PubMed Central - PubMed

Affiliation: Institute of Molecules and Materials, Radboud University Nijmegen, Nijmegen, The Netherlands.

ABSTRACT
Several reader domain proteins that specifically recognize methyllysine-containing histones contain the negatively-charged aspartate or glutamate residues as part of the aromatic cage. Herein, we report thermodynamic analyses for the recognition of histone H3K4me3 and H3K4me2 by the tandem tudor domain of Sgf29 and its recognition site variants. Small uncharged and large aromatic substitutions on the Asp266 site resulted in a significant decrease in binding affinities for both H3K4me3 and H3K4me2, demonstrating the role of the negative charge of Asp266 in the readout process by Sgf29. This study emphasizes the essential contribution of electrostatic interactions to the overall binding affinity, and reveals that the underlying mechanisms for the recognition of Kme2/3 depend on the composition and arrangement of the aromatic cage.

No MeSH data available.