Assessing the Delivery of Molecules to the Mitochondrial Matrix Using Click Chemistry

作者：Hoogewijs Kurt; James Andrew M; Smith Robin A J; Gait Michael J^*; Murphy Michael P^*; Lightowlers Robert N^*

来源：ChemBioChem, 2016, 17(14): 1312-1316.

DOI：10.1002/cbic.201600188

• click chemistry
• drug delivery
• mitochondria
• mitochondrial DNA
• targeting

Mitochondria are central to health and disease, hence there is considerable interest in developing mitochondria-targeted therapies that require the delivery of peptides or nucleic acid oligomers. However, progress has been impeded by the lack of a measure of mitochondrial import of these molecules. Here, we address this need by quantitatively detecting molecules within the mitochondrial matrix. We used a mitochondria-targeted cyclooctyne (MitoOct) that accumulates several-hundredfold in the matrix, driven by the membrane potential. There, MitoOct reacts through click chemistry with an azide on the target molecule to form a diagnostic product that can be quantified by mass spectrometry. Because the membrane potential-dependent MitoOct concentration in the matrix is essential for conjugation, we can now determine definitively whether a putative mitochondrion-targeted molecule reaches the matrix. This "ClickIn" approach will facilitate development of mitochondria-targeted therapies.

• 出版日期2016-7-15