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Cation-specific effects on enzymatic catalysis driven by interactions at the tunnel mouth

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Cation-specific effects on enzymatic catalysis driven by interactions at the tunnel mouth. / Štěpánková, Veronika; Paterová, Jana; Damborský, Jiří; Jungwirth, Pavel; Chaloupková, Radka; Heyda, Jan.

In: Journal of Physical Chemistry Part B, Vol. 117, No. 21, 30.05.2013, p. 6394-6402.

Research output: Contribution to journalArticleScientificpeer-review

Harvard

Štěpánková, V, Paterová, J, Damborský, J, Jungwirth, P, Chaloupková, R & Heyda, J 2013, 'Cation-specific effects on enzymatic catalysis driven by interactions at the tunnel mouth', Journal of Physical Chemistry Part B, vol. 117, no. 21, pp. 6394-6402. https://doi.org/10.1021/jp401506v

APA

Štěpánková, V., Paterová, J., Damborský, J., Jungwirth, P., Chaloupková, R., & Heyda, J. (2013). Cation-specific effects on enzymatic catalysis driven by interactions at the tunnel mouth. Journal of Physical Chemistry Part B, 117(21), 6394-6402. https://doi.org/10.1021/jp401506v

Vancouver

Štěpánková V, Paterová J, Damborský J, Jungwirth P, Chaloupková R, Heyda J. Cation-specific effects on enzymatic catalysis driven by interactions at the tunnel mouth. Journal of Physical Chemistry Part B. 2013 May 30;117(21):6394-6402. https://doi.org/10.1021/jp401506v

Author

Štěpánková, Veronika ; Paterová, Jana ; Damborský, Jiří ; Jungwirth, Pavel ; Chaloupková, Radka ; Heyda, Jan. / Cation-specific effects on enzymatic catalysis driven by interactions at the tunnel mouth. In: Journal of Physical Chemistry Part B. 2013 ; Vol. 117, No. 21. pp. 6394-6402.

Bibtex - Download

@article{a0db8fe6ebff42a5a057acdd249fdfad,
title = "Cation-specific effects on enzymatic catalysis driven by interactions at the tunnel mouth",
abstract = "Cationic specificity which follows the Hofmeister series has been established for the catalytic efficiency of haloalkane dehalogenase LinB by a combination of molecular dynamics simulations and enzyme kinetic experiments. Simulations provided a detailed molecular picture of cation interactions with negatively charged residues on the protein surface, particularly at the tunnel mouth leading to the enzyme active site. On the basis of the binding affinities, cations were ordered as Na+ > K+ > Rb+ > Cs+. In agreement with this result, a steady-state kinetic analysis disclosed that the smaller alkali cations influence formation and productivity of enzyme-substrate complexes more efficiently than the larger ones. A subsequent systematic investigation of two LinB mutants with engineered charge in the cation-binding site revealed that the observed cation affinities are enhanced by increasing the number of negatively charged residues at the tunnel mouth, and vice versa, reduced by decreasing this number. However, the cation-specific effects are overwhelmed by strong electrostatic interactions in the former case. Interestingly, the substrate inhibition of the mutant LinB L177D in the presence of chloride salts was 7 times lower than that of LinB wild type in glycine buffer. Our work provides new insight into the mechanisms of specific cation effects on enzyme activity and suggests a potential strategy for suppression of substrate inhibition by the combination of protein and medium engineering.",
author = "Veronika Štěp{\'a}nkov{\'a} and Jana Paterov{\'a} and Jiř{\'i} Damborsk{\'y} and Pavel Jungwirth and Radka Chaloupkov{\'a} and Jan Heyda",
year = "2013",
month = "5",
day = "30",
doi = "10.1021/jp401506v",
language = "English",
volume = "117",
pages = "6394--6402",
journal = "Journal of Physical Chemistry Part B",
issn = "1520-6106",
publisher = "American Chemical Society",
number = "21",

}

RIS (suitable for import to EndNote) - Download

TY - JOUR

T1 - Cation-specific effects on enzymatic catalysis driven by interactions at the tunnel mouth

AU - Štěpánková, Veronika

AU - Paterová, Jana

AU - Damborský, Jiří

AU - Jungwirth, Pavel

AU - Chaloupková, Radka

AU - Heyda, Jan

PY - 2013/5/30

Y1 - 2013/5/30

N2 - Cationic specificity which follows the Hofmeister series has been established for the catalytic efficiency of haloalkane dehalogenase LinB by a combination of molecular dynamics simulations and enzyme kinetic experiments. Simulations provided a detailed molecular picture of cation interactions with negatively charged residues on the protein surface, particularly at the tunnel mouth leading to the enzyme active site. On the basis of the binding affinities, cations were ordered as Na+ > K+ > Rb+ > Cs+. In agreement with this result, a steady-state kinetic analysis disclosed that the smaller alkali cations influence formation and productivity of enzyme-substrate complexes more efficiently than the larger ones. A subsequent systematic investigation of two LinB mutants with engineered charge in the cation-binding site revealed that the observed cation affinities are enhanced by increasing the number of negatively charged residues at the tunnel mouth, and vice versa, reduced by decreasing this number. However, the cation-specific effects are overwhelmed by strong electrostatic interactions in the former case. Interestingly, the substrate inhibition of the mutant LinB L177D in the presence of chloride salts was 7 times lower than that of LinB wild type in glycine buffer. Our work provides new insight into the mechanisms of specific cation effects on enzyme activity and suggests a potential strategy for suppression of substrate inhibition by the combination of protein and medium engineering.

AB - Cationic specificity which follows the Hofmeister series has been established for the catalytic efficiency of haloalkane dehalogenase LinB by a combination of molecular dynamics simulations and enzyme kinetic experiments. Simulations provided a detailed molecular picture of cation interactions with negatively charged residues on the protein surface, particularly at the tunnel mouth leading to the enzyme active site. On the basis of the binding affinities, cations were ordered as Na+ > K+ > Rb+ > Cs+. In agreement with this result, a steady-state kinetic analysis disclosed that the smaller alkali cations influence formation and productivity of enzyme-substrate complexes more efficiently than the larger ones. A subsequent systematic investigation of two LinB mutants with engineered charge in the cation-binding site revealed that the observed cation affinities are enhanced by increasing the number of negatively charged residues at the tunnel mouth, and vice versa, reduced by decreasing this number. However, the cation-specific effects are overwhelmed by strong electrostatic interactions in the former case. Interestingly, the substrate inhibition of the mutant LinB L177D in the presence of chloride salts was 7 times lower than that of LinB wild type in glycine buffer. Our work provides new insight into the mechanisms of specific cation effects on enzyme activity and suggests a potential strategy for suppression of substrate inhibition by the combination of protein and medium engineering.

UR - http://www.scopus.com/inward/record.url?scp=84878363659&partnerID=8YFLogxK

U2 - 10.1021/jp401506v

DO - 10.1021/jp401506v

M3 - Article

VL - 117

SP - 6394

EP - 6402

JO - Journal of Physical Chemistry Part B

JF - Journal of Physical Chemistry Part B

SN - 1520-6106

IS - 21

ER -