Abstract
Viability of isolated islets is one of the main obstacles limiting islet transplantation success. It has been reported that overexpression of Bcl-2/Bcl-XL proteins enhances islet viability. To avoid potential complications associated with long-term expression of anti-apoptotic proteins, we investigated the possibility of delivering Bcl-XL or its anti-apoptotic domain BH4 to islets by protein transduction. Bcl-XL and BH4 molecules were fused to TAT/PTD, the 11-aa cell penetrating peptide from HIV-1 transactivating protein, generating TAT-Bcl-XL and TAT-BH4, respectively. Transduction efficiency was assessed by laser scanning confocal microscopy of live islets. Biological activity was tested as the ability to protect NIT-1 insulinoma cell line from death induced by staurosporine or serum deprivation. Spontaneous caspase activation in human islets and cytotoxicity caused by IL-1β were significantly reduced in the presence of TAT-Bcl-XL and TAT-BH4. We conclude that both TAT proteins are biologically active after transduction and could be an asset in the improvement of islet viability.
Original language | English (US) |
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Pages (from-to) | 473-478 |
Number of pages | 6 |
Journal | Biochemical and biophysical research communications |
Volume | 323 |
Issue number | 2 |
DOIs | |
State | Published - Oct 15 2004 |
Keywords
- β-cells
- Bcl-XL
- BH4
- Islets
- Protein transduction
- Protein transduction domains
- TAT-PTD
ASJC Scopus subject areas
- Biochemistry
- Biophysics
- Molecular Biology