Import of TAT-Conjugated Propionyl Coenzyme A Carboxylase Using Models of Propionic Acidemia

2018 Collard TAT-PCC enzyme replacement mitochondrial import propionic acidemia.pdf (1.74 MB)

journal contribution

posted on 2019-03-08, 22:50 authored by Renata Collard, Tomas Majtan, Insun Park, Jan P. Kraus

Propionic acidemia is caused by a deficiency of the enzyme propionyl

coenzyme A carboxylase (PCC) located in the mitochondrial matrix. Cell-penetrating

peptides, including transactivator of transcription (TAT), offer a potential to deliver a

cargo into the mitochondrion. Here, we investigated the delivery of an 66 PCC enzyme

into mitochondria using the HIV TAT peptide at several levels: into isolated

mitochondria, in patient fibroblast cells, and in a mouse model. Results from Western

blots and enzyme activity assays confirmed the import of TAT-PCC into mitochondria,

as well as into patient fibroblasts, where the colocalization of imported

TAT-PCC and mitochondria was also confirmed by confocal fluorescence microscopy.

Furthermore, a single-dose intraperitoneal injection into PCC-deficient mice decreased

the propionylcarnitine/acetylcarnitine (C3/C2) ratio toward the normal level.

These results show that a cell-penetrating peptide can deliver active multimeric enzyme

into mitochondria in vitro, in situ, and in vivo and push the size limit of intracellular

delivery achieved so far. Our results are promising for other mitochondrionspecific

deficiencies.

History

16SDG30040000