Inhibition of type IA topoisomerase by a monoclonal antibody through perturbation of DNA cleavage-religation equilibrium

Majety Naga Leelaram, Anuradha Gopal Bhat, Shivanand Manjunath Hegde, Ramanathapuram Manjunath, Valakunja Nagaraja

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Type IA DNA topoisomerases, typically found in bacteria, are essential enzymes that catalyse the DNA relaxation of negative supercoils. DNA gyrase is the only type II topoisomerase that can carry out the opposite reaction (i.e. the introduction of the DNA supercoils). A number of diverse molecules target DNA gyrase. However, inhibitors that arrest the activity of bacterial topoisomerase I at low concentrations remain to be identified. Towards this end, as a proof of principle, monoclonal antibodies that inhibit Mycobacterium smegmatis topoisomerase I have been characterized and the specific inhibition of Mycobacterium smegmatis topoisomerase I by a monoclonal antibody, 2F3G4, at a nanomolar concentration is described. The enzyme-bound monoclonal antibody stimulated the first transesterification reaction leading to enhanced DNA cleavage, without significantly altering the religation activity of the enzyme. The stimulated DNA cleavage resulted in perturbation of the cleavage-religation equilibrium, increasing single-strand nicks and protein-DNA covalent adducts. Monoclonal antibodies with such a mechanism of inhibition can serve as invaluable tools for probing the structure and mechanism of the enzyme, as well as in the design of novel inhibitors that arrest enzyme activity.

Original languageEnglish (US)
Pages (from-to)55-65
Number of pages11
JournalFEBS Journal
Volume279
Issue number1
DOIs
StatePublished - Jan 2012
Externally publishedYes

Fingerprint

DNA Cleavage
Type I DNA Topoisomerase
Monoclonal Antibodies
DNA Gyrase
Mycobacterium smegmatis
DNA
Enzymes
DNA Topoisomerases
Type II DNA Topoisomerase
DNA Adducts
Enzyme Inhibitors
Transesterification
Enzyme activity
Bacteria
Molecules
Proteins

Keywords

  • monoclonal antibody
  • mycobacteria
  • protein-DNA complex
  • topoisomerase I
  • transesterification

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Inhibition of type IA topoisomerase by a monoclonal antibody through perturbation of DNA cleavage-religation equilibrium. / Leelaram, Majety Naga; Bhat, Anuradha Gopal; Hegde, Shivanand Manjunath; Manjunath, Ramanathapuram; Nagaraja, Valakunja.

In: FEBS Journal, Vol. 279, No. 1, 01.2012, p. 55-65.

Research output: Contribution to journalArticle

Leelaram, Majety Naga ; Bhat, Anuradha Gopal ; Hegde, Shivanand Manjunath ; Manjunath, Ramanathapuram ; Nagaraja, Valakunja. / Inhibition of type IA topoisomerase by a monoclonal antibody through perturbation of DNA cleavage-religation equilibrium. In: FEBS Journal. 2012 ; Vol. 279, No. 1. pp. 55-65.
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