Charge Interactions in a Highly Charge-Depleted Protein
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Charge Interactions in a Highly Charge-Depleted Protein. / Hervø-Hansen, Stefan; Højgaard, Casper; Johansson, Kristoffer Enøe; Wang, Yong; Wahni, Khadija; Young, David; Messens, Joris; Teilum, Kaare; Lindorff-Larsen, Kresten; Winther, Jakob Rahr.
In: Journal of the American Chemical Society, Vol. 143, No. 6, 2021, p. 2500-2508.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Charge Interactions in a Highly Charge-Depleted Protein
AU - Hervø-Hansen, Stefan
AU - Højgaard, Casper
AU - Johansson, Kristoffer Enøe
AU - Wang, Yong
AU - Wahni, Khadija
AU - Young, David
AU - Messens, Joris
AU - Teilum, Kaare
AU - Lindorff-Larsen, Kresten
AU - Winther, Jakob Rahr
PY - 2021
Y1 - 2021
N2 - Electrostatic forces are important for protein folding and are favored targets of protein engineering. However, interactions between charged residues are difficult to study because of the complex network of interactions found in most proteins. We have designed a purposely simple system to investigate this problem by systematically introducing individual and pairs of charged and titratable residues in a protein otherwise free of such residues. We used constant pH molecular dynamics simulations, NMR spectroscopy, and thermodynamic double mutant cycles to probe the structure and energetics of the interaction between the charged residues. We found that the partial burial of surface charges contributes to a shift in pKa value, causing an aspartate to titrate in the neutral pH range. Additionally, the interaction between pairs of residues was found to be highly context dependent, with some pairs having no apparent preferential interaction, while other pairs would engage in coupled titration forming a highly stabilized salt bridge. We find good agreement between experiments and simulations and use the simulations to rationalize our observations and to provide a detailed mechanistic understanding of the electrostatic interactions.
AB - Electrostatic forces are important for protein folding and are favored targets of protein engineering. However, interactions between charged residues are difficult to study because of the complex network of interactions found in most proteins. We have designed a purposely simple system to investigate this problem by systematically introducing individual and pairs of charged and titratable residues in a protein otherwise free of such residues. We used constant pH molecular dynamics simulations, NMR spectroscopy, and thermodynamic double mutant cycles to probe the structure and energetics of the interaction between the charged residues. We found that the partial burial of surface charges contributes to a shift in pKa value, causing an aspartate to titrate in the neutral pH range. Additionally, the interaction between pairs of residues was found to be highly context dependent, with some pairs having no apparent preferential interaction, while other pairs would engage in coupled titration forming a highly stabilized salt bridge. We find good agreement between experiments and simulations and use the simulations to rationalize our observations and to provide a detailed mechanistic understanding of the electrostatic interactions.
U2 - 10.1021/jacs.0c10789
DO - 10.1021/jacs.0c10789
M3 - Journal article
C2 - 33529004
VL - 143
SP - 2500
EP - 2508
JO - Journal of the American Chemical Society
JF - Journal of the American Chemical Society
SN - 0002-7863
IS - 6
ER -
ID: 256475367