Analysis of Etoposide Binding to Subdomains of Human DNA Topoisomerase IIα in the Absence of DNA

Article Article scientifique Article Analysis of Etoposide Binding to Subdomains of Human DNA Topoisomerase IIα in the Absence of DNA eng 2001 author Leroy Didier 134899 author Kajava Andrey V. author Frei Christian author Gasser Susan Margaret 133662 1624-1634 10.1021/bi0019141 0006-2960 https://pubs.acs.org/doi/10.1021/bi0019141 Epipodophyllotoxins are effective anti-tumor drugs that inhibit eukaryotic DNA topoisomerase II by trapping the enzyme in a covalent complex with DNA. We show that both the recombinant N-terminal ATPase domain and the B‘A‘ core domain of human topoisomerase IIα (htopoIIα) bind radiolabeled etoposide specifically, even in the absence of DNA. The addition of ATP impairs etoposide binding to the holoenzyme and the N-terminal domain, but not to the core domain. To see if this interference resembles that between novobiocin and ATP in the bacterial GyrB subunit, we modeled the structure of the N-terminal domain of htopoIIα and performed molecular docking analysis with etoposide. Mutagenesis of critical amino acids, predicted to stabilize the drug within the N-terminal domain, reveals a less efficient binding of etoposide to the mutated proteins as monitored by direct drug binding assays, although the binding of ATP is not affected. true Biochemistry 40 6 9c15a526-79e5-42c5-b890-046b36e1135c Article (Published version) Restricted bi0019141.pdf 1045222 application/pdf restrictedAccess

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Analysis of Etoposide Binding to Subdomains of Human DNA Topoisomerase IIα in the Absence of DNA	Biochemistry			2001	269	0

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Leroy, Didier