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ClickIn: A flexible protocol for quantifying mitochondrial uptake of nucleobase derivatives

Lookup NU author(s): Dr Kurt Hoogewijs, Professor Robert Lightowlers

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This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).

Abstract

© 2017 The Authors. There is an increasing interest in targeting molecules to the mitochondrial matrix. Many proteins are naturally imported through the translocase complexes found in the outer and inner mitochondrial membranes. One possible means for importing molecules is therefore to use a mitochondrial pre-protein as a vector and assess what forms of molecules can be attached to the pre-protein as cargo. A major difficulty with this approach is to ensure that any chimaeric molecule does indeed access the mitochondrial matrix and does not merely associate with the mitochondrial membranes. We have recently demonstrated that click chemistry can be used both to demonstrate convincingly mitochondrial import of a peptide–peptide nucleic acid conjugate and also to quantify the mitochondrial uptake for specific synthetic conjugates. We now report an adaptation of the synthesis to facilitate simple quantification of multiple molecules and hence to calculate the efficiency of their mitochondrial import.

Publication metadata

Author(s): Hoogewijs K, James AM, Smith RAJ, Abendroth F, Gait MJ, Murphy MP, Lightowlers RN

Publication type: Article

Publication status: Published

Journal: Interface Focus

Year: 2017

Volume: 7

Issue: 2

Online publication date: 17/02/2017

Acceptance date: 02/04/2016

Date deposited: 11/04/2017

ISSN (print): 2042-8898

ISSN (electronic): 2042-8901

Publisher: Royal Society

URL: https://doi.org/10.1098/rsfs.2016.0117

DOI: 10.1098/rsfs.2016.0117

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