FRET imaging of Pit-l protein interactions in living cells

Ammasi Periasamy, Richard Day

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

The combined use of fluorescence resonance energy transfer (FRET) microscopy and expression of genetic vectors encoding protein fusions with green fluorescent protein (GFP) and blue fluorescent protein (BFP) provides an exceptionally sensitive method for detecting the interaction of protein partners in living cells. The acquisition of FRET signals from GFP- and BFP-fusion proteins expressed in living cells was demonstrated using an optimized imaging system and high sensitivity charge coupled device camera. This imaging system was used to detect energy transfer signals from a fusion protein containing GFP physically linked to BFP expressed in living HeLa cells. In contrast, the co-localization of noninteracting GFP- and BFP-fusion proteins was not sufficient for energy transfer. The FRET imaging system was then used to demonstrate dimerization of the pituitary-specific transcription factor Pit-1 within the living cell nucleus.

Original languageEnglish (US)
Pages (from-to)154-160
Number of pages7
JournalJournal of Biomedical Optics
Volume3
Issue number2
StatePublished - 1998
Externally publishedYes

Fingerprint

Fluorescence Resonance Energy Transfer
resonance fluorescence
energy transfer
Cells
proteins
Proteins
Imaging techniques
Green Fluorescent Proteins
interactions
Fusion reactions
Imaging systems
Energy Transfer
fusion
Energy transfer
Transcription Factor Pit-1
Genetic Vectors
Dimerization
CCD cameras
Cell Nucleus
HeLa Cells

Keywords

  • Fluorescence resonance energy transfer (FRET)
  • Microscopy, green fluorescent proteins (GFPs)
  • Pit-1

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Radiology Nuclear Medicine and imaging
  • Atomic and Molecular Physics, and Optics
  • Radiological and Ultrasound Technology

Cite this

FRET imaging of Pit-l protein interactions in living cells. / Periasamy, Ammasi; Day, Richard.

In: Journal of Biomedical Optics, Vol. 3, No. 2, 1998, p. 154-160.

Research output: Contribution to journalArticle

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