Toggle Main Menu Toggle Search

Open Access padlockePrints

Specific targeting of a DNA-alkylating reagent to mitochondria: Synthesis and characterization of [4-((11aS)-7-methoxy-1,2,3,11a-tetrahydro-5H-pyrrolo[2,1-c][1,4] benzodiazepin-5-on-8-oxy)butyl]-triphenylphosphonium iodide

Lookup NU author(s): Professor Robert Lightowlers, Dr Paul Smith

Downloads

Full text for this publication is not currently held within this repository. Alternative links are provided below where available.


Abstract

The selective manipulation of the expression and replication of mitochondrial DNA (mtDNA) within mammalian cells has proven difficult. In progressing towards this goal we synthesized a novel mitochondria-targeted DNA-alkylating reagent. The active alkylating moiety [(11aS)-8-hydroxy-7-methoxy- 1,2,3,11 a-tetrahydro-5H-pyrrolo[2,1-c] [1,4]benzodiazepin-5-one (DC-81)], irreversibly alkylates guanine bases in DNA (with a preference for AGA triplets), preventing its expression and replication. To target this compound to mitochondria it was covalently coupled to the lipophilic triphenylphosphonium (TPP) cation to form a derivative referred to as mitoDC-81. Incorporation of this lipophilic cation led to the rapid uptake of mitoDC-81 by mitochondria, driven by the large membrane potential across the inner membrane. This compound efficiently alkylated isolated supercoiled, relaxed-circular or linear plasmid DNA and isolated mtDNA. However mitoDC-81 did not alkylate mtDNA within isolated mitochondria or cells, even though it accessed the mitochondrial matrix at concentrations up to 100-fold higher than those required to alkylate isolated DNA. This surprising finding suggests that mtDNA within intact mitochondria may not be accessible to this class of alkylating reagent. This inability to alkylate mtDNA in situ has significant implications for the design of therapies for mtDNA diseases and for studies on the packaging, expression and turnover of mtDNA in general.


Publication metadata

Author(s): James AM, Blaikie FH, Smith RAJ, Lightowlers RN, Smith PM, Murphy MP

Publication type: Article

Publication status: Published

Journal: European Journal of Biochemistry

Year: 2003

Volume: 270

Issue: 13

Pages: 2827-2836

ISSN (print): 0014-2956

ISSN (electronic):

Publisher: Wiley-Blackwell

URL: http://dx.doi.org/10.1046/j.1432-1033.2003.03660.x

DOI: 10.1046/j.1432-1033.2003.03660.x

PubMed id: 12823553


Altmetrics

Altmetrics provided by Altmetric


Actions

Find at Newcastle University icon    Link to this publication


Share