TAT-Conjugated NDUFS8 Can Be Transduced into Mitochondria in a Membrane-Potential-Independent Manner and Rescue Complex I Deficiency

Autor: Chao-Chang Lee, Bo-Yu Lin, Pin-Chao Liao, Kai-Wen Teng, Juan-Yu Chang, Gui-Teng Zheng, Mou-Chieh Kao
Jazyk: angličtina
Rok vydání: 2021
Předmět:
0301 basic medicine
HIV-1 transactivator of transcription peptide (TAT)
NDUFS8
viruses
Mitochondrion
protein transduction domain
Transduction (genetics)
RNA
Small Interfering
Biology (General)
Cells
Cultured
Spectroscopy
Membrane Potential
Mitochondrial
biology
Chemistry
complex I
NADH dehydrogenase
General Medicine
Transfection
Computer Science Applications
Cell biology
mitochondria
Protein Transport
mitochondrial targeting sequence
RNA Interference
tat Gene Products
Human Immunodeficiency Virus
enzyme replacement therapy
Cell Survival
QH301-705.5
Recombinant Fusion Proteins
Protein subunit
Catalysis
Article
Cell Line
Inorganic Chemistry
03 medical and health sciences
mitochondrial membrane potential
Humans
Amino Acid Sequence
RNA
Messenger
Physical and Theoretical Chemistry
QD1-999
Molecular Biology
Electron Transport Complex I
030102 biochemistry & molecular biology
Organic Chemistry
NADH Dehydrogenase
Fusion protein
030104 developmental biology
Electron Transport Chain Complex Proteins
Cell culture
biology.protein
Zdroj: International Journal of Molecular Sciences
Volume 22
Issue 12
International Journal of Molecular Sciences, Vol 22, Iss 6524, p 6524 (2021)
ISSN: 1422-0067
DOI: 10.3390/ijms22126524
Popis: NADH dehydrogenase (ubiquinone) Fe-S protein 8 (NDUFS8) is a nuclear-encoded core subunit of human mitochondrial complex I. Defects in NDUFS8 are associated with Leigh syndrome and encephalomyopathy. Cell-penetrating peptide derived from the HIV-1 transactivator of transcription protein (TAT) has been successfully applied as a carrier to bring fusion proteins into cells without compromising the biological function of the cargoes. In this study, we developed a TAT-mediated protein transduction system to rescue complex I deficiency caused by NDUFS8 defects. Two fusion proteins (TAT-NDUFS8 and NDUFS8-TAT) were exogenously expressed and purified from Escherichia coli for transduction of human cells. In addition, similar constructs were generated and used in transfection studies for comparison. The results showed that both exogenous TAT-NDUFS8 and NDUFS8-TAT were delivered into mitochondria and correctly processed. Interestingly, the mitochondrial import of TAT-containing NDUFS8 was independent of mitochondrial membrane potential. Treatment with TAT-NDUFS8 not only significantly improved the assembly of complex I in an NDUFS8-deficient cell line, but also partially rescued complex I functions both in the in-gel activity assay and the oxygen consumption assay. Our current findings suggest the considerable potential of applying the TAT-mediated protein transduction system for treatment of complex I deficiency.
Databáze: OpenAIRE
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