Abstract
In this study we provide in vitro and in vivo evidence showing that the protein disulphide isomerase (PDI) activity of type 2 transglutaminase (TG2) regulates the correct assembly and function of the mitochondrial ADP/ATP transporter adenine nucleotide translocator 1 (ANT1). We demonstrate, by means of biochemical and morphological analyses, that ANT1 and TG2 physically interact in the mitochondria. Under physiological conditions, TG2's PDI activity regulates the ADP/ATP transporter function by controlling the oligomerization of ANT1. In fact, mitochondria isolated from hearts of TG2−/− mice exhibit increased polymerization of ANT1, paralleled by an enhanced ADP/ATP carrier activity, as compared to mitochondria belonging to TG2+/+ mice. Interestingly, upon cell-death induction, ANT1 becomes a substrate for TG2's cross-linking activity and the lack of TG2 results in a reduction of apoptosis as well as in a marked sensitivity to the ADP/ATP exchange inhibition by atractyloside. These findings suggest a complex TG2-dependent regulation of the ADP/ATP transporter and reveal new important avenues for its potential applications in the treatment of some mitochondrial-dependent diseases, including cardiovascular and neurodegenerative diseases.
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Abbreviations
- ANT1:
-
adenine nucleotide translocator 1
- ATRL:
-
atractyloside
- BA:
-
bongkrekic acid
- CyA:
-
cyclosporine A
- CyP-D:
-
cyclophilin D
- DDG:
-
deoxy-D-glucose
- EZ-link:
-
5-(biotinamido)pentylamine
- IMM:
-
inner mitochondrial membrane
- IVM:
-
intensified video microscopy
- JC-1:
-
5-5′,6-6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazol-carbocyanine iodide
- MPP+:
-
1-methyl-4-phenyl-pyridinium ion
- K/S test:
-
Kolmogorov–Smirnov test
- Δψm:
-
mitochondrial membrane potential
- MMP:
-
mitochondrial membrane potential
- MAb:
-
monoclonal antibody
- MODY:
-
maturity-onset diabetes of the young
- MEF:
-
mouse embryonic fibroblast
- 3-NP:
-
3-nitropropionic acid
- OXPHOS:
-
oxidative phosphorylation
- PAb:
-
polyclonal antibody
- PDI:
-
protein disulphide isomerase
- PI:
-
presidium iodide
- PLC δ1:
-
phospholipase C
- ROS:
-
reactive oxygen species
- S.D.:
-
standard deviation
- STS:
-
staurosporine
- TG2:
-
type 2 transglutaminase
- TMRM:
-
tetramethylrhodamine ester
- VDAC:
-
voltage-dependent anion channel
- WCE:
-
whole cell extracts
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Acknowledgements
We thank Vito Porcelli for skilled technical assistance. This work was supported by grants from: Prin 2006 to MP and CR; Associazione Italiana Ricerca sul Cancro to MP; Telethon (GGP06254) to MP and WM; Ministero della Sanità to WM; EU-FP6 (LSHM-CT-2004-503116) to LP.
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Malorni, W., Farrace, M., Matarrese, P. et al. The adenine nucleotide translocator 1 acts as a type 2 transglutaminase substrate: implications for mitochondrial-dependent apoptosis. Cell Death Differ 16, 1480–1492 (2009). https://doi.org/10.1038/cdd.2009.100
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DOI: https://doi.org/10.1038/cdd.2009.100
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