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 |
|---|---|
| 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 |
Fingerprint
Keywords
- β-cells
- Bcl-XL
- BH4
- Islets
- Protein transduction
- Protein transduction domains
- TAT-PTD
ASJC Scopus subject areas
- Biochemistry
- Biophysics
- Molecular Biology
Cite this
Delivery of Bcl-XL or its BH4 domain by protein transduction inhibits apoptosis in human islets. / Klein, Dagmar; Ribeiro, Melina M.; Mendoza, Valeska; Jayaraman, Sundararajan; Kenyon, Norma S; Pileggi, Antonello; Molano, Ruth; Inverardi, Luca A; Ricordi, Camillo; Pastori, Ricardo.
In: Biochemical and Biophysical Research Communications, Vol. 323, No. 2, 15.10.2004, p. 473-478.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Delivery of Bcl-XL or its BH4 domain by protein transduction inhibits apoptosis in human islets
AU - Klein, Dagmar
AU - Ribeiro, Melina M.
AU - Mendoza, Valeska
AU - Jayaraman, Sundararajan
AU - Kenyon, Norma S
AU - Pileggi, Antonello
AU - Molano, Ruth
AU - Inverardi, Luca A
AU - Ricordi, Camillo
AU - Pastori, Ricardo
PY - 2004/10/15
Y1 - 2004/10/15
N2 - 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.
AB - 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.
KW - β-cells
KW - Bcl-XL
KW - BH4
KW - Islets
KW - Protein transduction
KW - Protein transduction domains
KW - TAT-PTD
UR - http://www.scopus.com/inward/record.url?scp=4544325016&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=4544325016&partnerID=8YFLogxK
U2 - 10.1016/j.bbrc.2004.08.116
DO - 10.1016/j.bbrc.2004.08.116
M3 - Article
C2 - 15369775
AN - SCOPUS:4544325016
VL - 323
SP - 473
EP - 478
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 2
ER -