Uncoupling conformational states from activity in an allosteric enzyme

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Uncoupling conformational states from activity in an allosteric enzyme. / Pisco, João P; de Chiara, Cesira; Pacholarz, Kamila J; Garza-Garcia, Acely; Ogrodowicz, Roksana W; Walker, Philip A; Barran, Perdita E; Smerdon, Stephen J; de Carvalho, Luiz Pedro S.

In: Nature Communications, Vol. 8, No. 1, 203, 07.08.2017.

Research output: Contribution to journalArticlepeer-review

Harvard

Pisco, JP, de Chiara, C, Pacholarz, KJ, Garza-Garcia, A, Ogrodowicz, RW, Walker, PA, Barran, PE, Smerdon, SJ & de Carvalho, LPS 2017, 'Uncoupling conformational states from activity in an allosteric enzyme', Nature Communications, vol. 8, no. 1, 203. https://doi.org/10.1038/s41467-017-00224-0

APA

Pisco, J. P., de Chiara, C., Pacholarz, K. J., Garza-Garcia, A., Ogrodowicz, R. W., Walker, P. A., Barran, P. E., Smerdon, S. J., & de Carvalho, L. P. S. (2017). Uncoupling conformational states from activity in an allosteric enzyme. Nature Communications, 8(1), [203]. https://doi.org/10.1038/s41467-017-00224-0

Vancouver

Pisco JP, de Chiara C, Pacholarz KJ, Garza-Garcia A, Ogrodowicz RW, Walker PA et al. Uncoupling conformational states from activity in an allosteric enzyme. Nature Communications. 2017 Aug 7;8(1). 203. https://doi.org/10.1038/s41467-017-00224-0

Author

Pisco, João P ; de Chiara, Cesira ; Pacholarz, Kamila J ; Garza-Garcia, Acely ; Ogrodowicz, Roksana W ; Walker, Philip A ; Barran, Perdita E ; Smerdon, Stephen J ; de Carvalho, Luiz Pedro S. / Uncoupling conformational states from activity in an allosteric enzyme. In: Nature Communications. 2017 ; Vol. 8, No. 1.

Bibtex

@article{6281d4efda574e7f9de180fc790b7a4f,
title = "Uncoupling conformational states from activity in an allosteric enzyme",
abstract = "ATP-phosphoribosyltransferase (ATP-PRT) is a hexameric enzyme in conformational equilibrium between an open and seemingly active state and a closed and presumably inhibited form. The structure-function relationship of allosteric regulation in this system is still not fully understood. Here, we develop a screening strategy for modulators of ATP-PRT and identify 3-(2-thienyl)-L-alanine (TIH) as an allosteric activator of this enzyme. Kinetic analysis reveals co-occupancy of the allosteric sites by TIH and L-histidine. Crystallographic and native ion-mobility mass spectrometry data show that the TIH-bound activated form of the enzyme closely resembles the inhibited L-histidine-bound closed conformation, revealing the uncoupling between ATP-PRT open and closed conformations and its functional state. These findings suggest that dynamic processes are responsible for ATP-PRT allosteric regulation and that similar mechanisms might also be found in other enzymes bearing a ferredoxin-like allosteric domain.Active and inactive state ATP-phosphoribosyltransferases (ATP-PRTs) are believed to have different conformations. Here the authors show that in both states, ATP-PRT has a similar structural arrangement, suggesting that dynamic alterations are involved in ATP-PRT regulation by allosteric modulators.",
keywords = "ATP Phosphoribosyltransferase/chemistry, Adenosine Triphosphate/chemistry, Allosteric Regulation, Allosteric Site, Histidine/chemistry, Kinetics, Models, Molecular",
author = "Pisco, {Jo{\~a}o P} and {de Chiara}, Cesira and Pacholarz, {Kamila J} and Acely Garza-Garcia and Ogrodowicz, {Roksana W} and Walker, {Philip A} and Barran, {Perdita E} and Smerdon, {Stephen J} and {de Carvalho}, {Luiz Pedro S}",
year = "2017",
month = aug,
day = "7",
doi = "10.1038/s41467-017-00224-0",
language = "English",
volume = "8",
journal = "Nature Communications",
issn = "2041-1723",
publisher = "Springer",
number = "1",

}

RIS

TY - JOUR

T1 - Uncoupling conformational states from activity in an allosteric enzyme

AU - Pisco, João P

AU - de Chiara, Cesira

AU - Pacholarz, Kamila J

AU - Garza-Garcia, Acely

AU - Ogrodowicz, Roksana W

AU - Walker, Philip A

AU - Barran, Perdita E

AU - Smerdon, Stephen J

AU - de Carvalho, Luiz Pedro S

PY - 2017/8/7

Y1 - 2017/8/7

N2 - ATP-phosphoribosyltransferase (ATP-PRT) is a hexameric enzyme in conformational equilibrium between an open and seemingly active state and a closed and presumably inhibited form. The structure-function relationship of allosteric regulation in this system is still not fully understood. Here, we develop a screening strategy for modulators of ATP-PRT and identify 3-(2-thienyl)-L-alanine (TIH) as an allosteric activator of this enzyme. Kinetic analysis reveals co-occupancy of the allosteric sites by TIH and L-histidine. Crystallographic and native ion-mobility mass spectrometry data show that the TIH-bound activated form of the enzyme closely resembles the inhibited L-histidine-bound closed conformation, revealing the uncoupling between ATP-PRT open and closed conformations and its functional state. These findings suggest that dynamic processes are responsible for ATP-PRT allosteric regulation and that similar mechanisms might also be found in other enzymes bearing a ferredoxin-like allosteric domain.Active and inactive state ATP-phosphoribosyltransferases (ATP-PRTs) are believed to have different conformations. Here the authors show that in both states, ATP-PRT has a similar structural arrangement, suggesting that dynamic alterations are involved in ATP-PRT regulation by allosteric modulators.

AB - ATP-phosphoribosyltransferase (ATP-PRT) is a hexameric enzyme in conformational equilibrium between an open and seemingly active state and a closed and presumably inhibited form. The structure-function relationship of allosteric regulation in this system is still not fully understood. Here, we develop a screening strategy for modulators of ATP-PRT and identify 3-(2-thienyl)-L-alanine (TIH) as an allosteric activator of this enzyme. Kinetic analysis reveals co-occupancy of the allosteric sites by TIH and L-histidine. Crystallographic and native ion-mobility mass spectrometry data show that the TIH-bound activated form of the enzyme closely resembles the inhibited L-histidine-bound closed conformation, revealing the uncoupling between ATP-PRT open and closed conformations and its functional state. These findings suggest that dynamic processes are responsible for ATP-PRT allosteric regulation and that similar mechanisms might also be found in other enzymes bearing a ferredoxin-like allosteric domain.Active and inactive state ATP-phosphoribosyltransferases (ATP-PRTs) are believed to have different conformations. Here the authors show that in both states, ATP-PRT has a similar structural arrangement, suggesting that dynamic alterations are involved in ATP-PRT regulation by allosteric modulators.

KW - ATP Phosphoribosyltransferase/chemistry

KW - Adenosine Triphosphate/chemistry

KW - Allosteric Regulation

KW - Allosteric Site

KW - Histidine/chemistry

KW - Kinetics

KW - Models, Molecular

U2 - 10.1038/s41467-017-00224-0

DO - 10.1038/s41467-017-00224-0

M3 - Article

C2 - 28781362

VL - 8

JO - Nature Communications

JF - Nature Communications

SN - 2041-1723

IS - 1

M1 - 203

ER -