Targeting proteins to mitochondria using TAT

Victoria Del Gaizo, James A. MacKenzie, R. Payne

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

Disorders of mitochondrial function cause significant human disease and suffering. To date, correction of these mitochondria defects has depended on biochemical approaches and has not been achieved via gene therapies. Using previously described fusion proteins containing the transactivator of transcription (TAT) region from the HIV virus and green fluorescent protein (GFP), with and without a mitochondrial targeting sequence (MTS) from mitochondrial malate dehydrogenase (mMDH), we have investigated transduction across mitochondrial membranes. Both TAT-GFP and TAT-mMDH-GFP fusion proteins are protected from externally added protease when incubated with isolated mitochondria. Furthermore, both TAT fusion proteins rapidly enter cultured cells and transduce into mitochondria. However, the MTS allows processing of the fusion protein and is necessary for persistence in mitochondria over time. Neither degradation of import receptors nor disruption of the mitochondrial membrane potential or pH gradient inhibits protein transduction of either fusion protein. Furthermore, when injected into pregnant mice, TAT-mMDH-GFP is detectable throughout fetal and neonatal pups. These results indicate that TAT fusion proteins are able to traverse mitochondrial membranes through mechanisms that do not involve the regular import pathway, and that the addition of a MTS allows persistence of the fusion protein within mitochondria. TAT-MTS fusion proteins may represent a viable option as potential mitochondrial protein therapies.

Original languageEnglish (US)
Pages (from-to)170-180
Number of pages11
JournalMolecular Genetics and Metabolism
Volume80
Issue number1-2
DOIs
StatePublished - Sep 2003
Externally publishedYes

Fingerprint

Mitochondria
Trans-Activators
Protein Transport
Transcription
Fusion reactions
Green Fluorescent Proteins
Malate Dehydrogenase
Proteins
Mitochondrial Membranes
Membranes
Gene therapy
Mitochondrial Diseases
Proton-Motive Force
Mitochondrial Membrane Potential
Mitochondrial Proteins
Viruses
Psychological Stress
Genetic Therapy
Cultured Cells
Peptide Hydrolases

Keywords

  • CMXRos
  • GFP
  • Mitochondria
  • Mitochondrial diseases
  • Mitochondrial malate dehydrogenase
  • Mitochondrial membrane potential
  • Placenta
  • Protein sorting signals
  • PTD
  • TAT

ASJC Scopus subject areas

  • Biochemistry
  • Genetics
  • Endocrinology, Diabetes and Metabolism

Cite this

Targeting proteins to mitochondria using TAT. / Del Gaizo, Victoria; MacKenzie, James A.; Payne, R.

In: Molecular Genetics and Metabolism, Vol. 80, No. 1-2, 09.2003, p. 170-180.

Research output: Contribution to journalArticle

Del Gaizo, Victoria ; MacKenzie, James A. ; Payne, R. / Targeting proteins to mitochondria using TAT. In: Molecular Genetics and Metabolism. 2003 ; Vol. 80, No. 1-2. pp. 170-180.
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