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Hidden Allostery in 14-3-3 Proteins Orchestrated by ATP
CC BY 4.0 · Indian J Med Paediatr Oncol 2024; 45(S 01): S1-S16
DOI: DOI: 10.1055/s-0044-1788229
*Corresponding author: (e-mail: vprasanna@actrec.gov.in).
Abstract
Background: 14-3-3 proteins are a family of highly conserved proteins that bind and regulate the functions of different phosphoproteins and some nonphosphoproteins. These proteins have also been credited with enzymatic activities. We had earlier reported that many human 14-3-3 isoforms can hydrolyze ATP. However, the catalytic residues involved and the significance of this enzymatic function remained unclear.
Materials and Methods: In this study we performed structure guided docking of 14-3-3 using Schrödinger software and limited-proteolysis coupled with mass spectroscopy to identify ATP binding site. We performed site-directed mutagenesis to confirm the sites. We navigated through the various peptide ligands of 14-3-3 to test the effect of ATP binding on inherent ligand binding property of 14-3-3.
Results: Molecular docking study and Lip-MS provided two putative ATP binding sites in 14-3-3. We checked these two sites and confirmed the sites by mutagenesis. Specifically, two glutamic acid residues are involved in the hydrolysis of ATP, probably aided by bound water. We found that the binding of one of the nonphosphopeptides was significantly affected by the presence of ATP and ATPγS but such an effect was not seen in any tested phosphopeptides.
Conclusion: These studies unveil the hidden allosteric properties of the 14-3-3 proteins and their role in excluding specific ligands from the binding pocket. Such selectivity may play an important context-specific regulation of 14-3-3 functions. We hereby propose that 14-3-3 is an unconventional ATPase lacking classical ATP binding motifs and folds, but executes catalysis via a well-known catalytic mechanism.
Publication History
Article published online:
08 July 2024
© 2024. The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. (https://creativecommons.org/licenses/by/4.0/)
Thieme Medical and Scientific Publishers Pvt. Ltd.
A-12, 2nd Floor, Sector 2, Noida-201301 UP, India
*Corresponding author: (e-mail: vprasanna@actrec.gov.in).
Abstract
Background: 14-3-3 proteins are a family of highly conserved proteins that bind and regulate the functions of different phosphoproteins and some nonphosphoproteins. These proteins have also been credited with enzymatic activities. We had earlier reported that many human 14-3-3 isoforms can hydrolyze ATP. However, the catalytic residues involved and the significance of this enzymatic function remained unclear.
Materials and Methods: In this study we performed structure guided docking of 14-3-3 using Schrödinger software and limited-proteolysis coupled with mass spectroscopy to identify ATP binding site. We performed site-directed mutagenesis to confirm the sites. We navigated through the various peptide ligands of 14-3-3 to test the effect of ATP binding on inherent ligand binding property of 14-3-3.
Results: Molecular docking study and Lip-MS provided two putative ATP binding sites in 14-3-3. We checked these two sites and confirmed the sites by mutagenesis. Specifically, two glutamic acid residues are involved in the hydrolysis of ATP, probably aided by bound water. We found that the binding of one of the nonphosphopeptides was significantly affected by the presence of ATP and ATPγS but such an effect was not seen in any tested phosphopeptides.
Conclusion: These studies unveil the hidden allosteric properties of the 14-3-3 proteins and their role in excluding specific ligands from the binding pocket. Such selectivity may play an important context-specific regulation of 14-3-3 functions. We hereby propose that 14-3-3 is an unconventional ATPase lacking classical ATP binding motifs and folds, but executes catalysis via a well-known catalytic mechanism.
No conflict of interest has been declared by the author(s).
Publication History
Article published online:
08 July 2024
© 2024. The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. (https://creativecommons.org/licenses/by/4.0/)
Thieme Medical and Scientific Publishers Pvt. Ltd.
A-12, 2nd Floor, Sector 2, Noida-201301 UP, India