Regulatory and Catalytic Domains of Poly(ADP-ribose) Polymerases Cross-Complement for DNA-Break-Dependent Allosteric Stimulation of Catalytic Activity DOI
Milan Makwana,

G. P.,

Waghela Deeksha

et al.

ACS Chemical Biology, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 11, 2025

Allosteric regulation is achieved by regulatory domains that sense stimuli and induce conformational changes in the functional domain performs catalytic activity of enzyme. Poly-ADP-ribose polymerases (PARPs) are modular enzymes present across all life including Archaea, Bacteria, Eukarya. A typical architecture PARPs consists a conserved C-terminal (CAT) associated with multiple distinct N-terminal sensory and/or which together serve as region (REG). In this study, we investigated whether REG different orthologs paralogs from mammals (hPARP1 hPARP2), plants (atPARP2), bacteria (haPARP) can assemble CAT each other to generate chimeric assemblies. We have employed qualitative quantitative enzyme assays along binding studies examine these vitro The cis-complemented hPARP2 exhibited micromolar affinity, suggesting interact independent allosteric ligands. Also, our results show PARP proteins functionally active conformation presence DNA implying not required be on single polypeptide for stimulation. Interestingly, only atPARP2 displayed complementation studied PARPs. Conversely, hPARP1 failed cross-complement while showed robust cross-complementation. Our novel assemblies developed powerful synthetic biology tool interrogate control their activities living cells. addition, co-engineering non-complementing PARPs, new selective ligand-dependent systems. Furthermore, study facilitate understanding coevolution enzymes.

Language: Английский

Regulatory and Catalytic Domains of Poly(ADP-ribose) Polymerases Cross-Complement for DNA-Break-Dependent Allosteric Stimulation of Catalytic Activity DOI
Milan Makwana,

G. P.,

Waghela Deeksha

et al.

ACS Chemical Biology, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 11, 2025

Allosteric regulation is achieved by regulatory domains that sense stimuli and induce conformational changes in the functional domain performs catalytic activity of enzyme. Poly-ADP-ribose polymerases (PARPs) are modular enzymes present across all life including Archaea, Bacteria, Eukarya. A typical architecture PARPs consists a conserved C-terminal (CAT) associated with multiple distinct N-terminal sensory and/or which together serve as region (REG). In this study, we investigated whether REG different orthologs paralogs from mammals (hPARP1 hPARP2), plants (atPARP2), bacteria (haPARP) can assemble CAT each other to generate chimeric assemblies. We have employed qualitative quantitative enzyme assays along binding studies examine these vitro The cis-complemented hPARP2 exhibited micromolar affinity, suggesting interact independent allosteric ligands. Also, our results show PARP proteins functionally active conformation presence DNA implying not required be on single polypeptide for stimulation. Interestingly, only atPARP2 displayed complementation studied PARPs. Conversely, hPARP1 failed cross-complement while showed robust cross-complementation. Our novel assemblies developed powerful synthetic biology tool interrogate control their activities living cells. addition, co-engineering non-complementing PARPs, new selective ligand-dependent systems. Furthermore, study facilitate understanding coevolution enzymes.

Language: Английский

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