Transduction of TAT fusion proteins into osteoclasts and osteoblasts

Svetlana Dolgilevich, Neeha Zaidi, Jianbo Song, Etsuko Abe, Baljit S. Moonga, Li Sun

Research output: Contribution to journalArticle

Abstract

It has been difficult to transduce primary cultures of bone cells with proteins of interest. Here, we report the development and validation of a new technology for transduction of osteoblasts and osteoclasts with peptides and moderately sized proteins. Fusion proteins between TAT, an 11 amino acid Arg-rich sequence derived from the HIV protein, and either hemagglutinin or calcineurin Aα were synthesized and purified. Exposure of osteoblasts and osteoclasts in primary culture to either TAT-HA or TAT-calcineurin Aα resulted in a rapid (within 10 min) intracellular movement of the fusion protein evident on co-immunostaining. Almost 99% of cells were transduced and the fusion protein was retained in ∼50% of the cells for up to 5 days. TAT did not abolish the functionality of calcineurin Aα; the fusion protein stimulated osteoblast differentiation and inhibited osteoclastic resorption. We expect that our studies will provide a firm basis for the future development of TAT fusion proteins for critical molecules involved in bone cell differentiation and function.

Original languageEnglish (US)
Pages (from-to)505-509
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume299
Issue number3
DOIs
StatePublished - Dec 2 2002
Externally publishedYes

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Osteoblasts
Osteoclasts
Fusion reactions
Calcineurin
Proteins
Cell culture
Bone
Human Immunodeficiency Virus Proteins
Bone and Bones
Primary Cell Culture
Hemagglutinins
Cell Differentiation
Technology
Amino Acids
Peptides
Molecules

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Transduction of TAT fusion proteins into osteoclasts and osteoblasts. / Dolgilevich, Svetlana; Zaidi, Neeha; Song, Jianbo; Abe, Etsuko; Moonga, Baljit S.; Sun, Li.

In: Biochemical and Biophysical Research Communications, Vol. 299, No. 3, 02.12.2002, p. 505-509.

Research output: Contribution to journalArticle

Dolgilevich, Svetlana ; Zaidi, Neeha ; Song, Jianbo ; Abe, Etsuko ; Moonga, Baljit S. ; Sun, Li. / Transduction of TAT fusion proteins into osteoclasts and osteoblasts. In: Biochemical and Biophysical Research Communications. 2002 ; Vol. 299, No. 3. pp. 505-509.
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