Delivery of Bcl-XL or its BH4 domain by protein transduction inhibits apoptosis in human islets

Dagmar Klein, Melina M. Ribeiro, Valeska Mendoza, Sundararajan Jayaraman, Norma S Kenyon, Antonello Pileggi, Ruth Molano, Luca A Inverardi, Camillo Ricordi, Ricardo Pastori

Research output: Contribution to journalArticle

66 Citations (Scopus)

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 languageEnglish
Pages (from-to)473-478
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume323
Issue number2
DOIs
StatePublished - Oct 15 2004

Fingerprint

Apoptosis
bcl-X Protein
Cell-Penetrating Peptides
Islets of Langerhans Transplantation
Insulinoma
Apoptosis Regulatory Proteins
Proteins
Staurosporine
Confocal microscopy
Cytotoxicity
Caspases
Bioactivity
Interleukin-1
Confocal Microscopy
HIV-1
Cell Death
Chemical activation
Cells
Scanning
Cell Line

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 journalArticle

Klein, Dagmar ; Ribeiro, Melina M. ; Mendoza, Valeska ; Jayaraman, Sundararajan ; Kenyon, Norma S ; Pileggi, Antonello ; Molano, Ruth ; Inverardi, Luca A ; Ricordi, Camillo ; Pastori, Ricardo. / Delivery of Bcl-XL or its BH4 domain by protein transduction inhibits apoptosis in human islets. In: Biochemical and Biophysical Research Communications. 2004 ; Vol. 323, No. 2. pp. 473-478.
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